Collection, Labelling, Handling and Care of Specimen - Part 2

In Partial Fulfillment of MC3: Microbiology and Parasitology Laboratory Course

Submitted by:
Nacua, Derick - Group Leader
Campaner, Zein
Carpena, Jermin
Colina, Rowenrey D.
Collins, Cassandra
Ediza, Louise Ysabel
Fernandez, Philip
Lao, Thea
Limboy, James Yap
Nacario, Xuanne Nina
Onil, Raymond Reave
Quiñones, Kristine Marie H.
Reyes, Lualhati
Tejada, Whel Marie S.

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Table of Contents

Title Page Number

Introduction 5

1.1 Urine 6

● Collection 7

● Package 13

● Transport 16

● Storage 18

1.2 Sputum 20

● Collection 20

● Package 28

● Transport 33

● Storage 33

1.3 Vaginal Fluid 35

● Collection 35

● Package 37

● Transport 38

● Storage 36

1.4 Pericardial Fluid 39

● Collection 40

● Package 41

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● Transport 41

● Storage 42

1.5 Synovial Fluid 43

● Collecting 43

● Packaging 49

● Transport 50

● Storage 51

1.6 Peritoneal Fluid 52

● Collecting 53

● Packaging 57

● Transport 57

● Storage 57

1.7 Seminal Fluid 58

● Collecting 58

● Packaging 61

● Transport 62

● Storage 63

1.8 Cerebrospinal Fluid 64

● Collecting 66

● Packaging 75

● Transport 66

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● Storage 76

1.9 Gastric Fluid 77

● Collecting 78

● Packaging 82

● Transport 89

● Storage 89

2.0 Pleural Fluid 90

● Collecting 91

● Packaging 92

● Transport 92

● Storage 93

References: 95

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INTRODUCTION

HUMAN BIOLOGICAL SPECIMEN

Any material derived from a human such as blood, urine, tissues, organs, saliva,

DNA/RNA, hair, nail clippings, or any other cells or fluids-whether collected for research

purposes or as residual specimens from diagnostic, therapeutic, or surgical procedures

is called a human biological specimen.

Human biological specimens are considered a biohazard and should be treated as

potentially infectious at all times. That being said, universal precautions are the best

practices to avoid accidental lab exposure to infectious agents in the collection,

packaging, storing, and transportation of such materials: 1. Personal protective

equipment such as gloves, lab coats, and eye protection must be worn appropriately at

all times. 2. Knowledge of safe practices with the needles, glass, and other sharps that

might have traces of human specimen on them. 3. Proper use of decontaminating

agents. Again, all human specimen is treated as potentially infectious, even the samples

that have been processed or frozen.

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1.1 Urine

PRINCIPLE

Urine has a long history of being used as a specimen in clinical labs for analysis.

Urine, after blood, is the most widely used specimen for diagnostic testing, disease

monitoring, and drug detection. Urine testing, both automated and manual processes,

is becoming more prominent. Urine specimen collection and transportation to the

clinical laboratory are critical because variables such as collection process, container,

transportation, and storage influence research outcome and, as a result, diagnostic and

therapeutic decisions based on the findings. Nurses or clinical staff are in charge of

clinical care, collecting and labeling urine specimens, and transporting specimens to the

Laboratory on time.

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COLLECTION OF URINE SPECIMEN

A. Specimen types

1. Random specimen

For chemical and microscopic examination, a voided specimen is usually more

suitable. A randomly collected specimen may be collected at unspecified times and is

often more convenient for the patient. A random specimen is suitable for most

screening purposes.

2. First morning specimen or 8-hour specimen

The patient should be instructed to collect the specimen immediately upon rising

from a night’s sleep. Other 8-hour periods may be used to accommodate insomniacs,

night-shift workers, and in certain pediatric situations. The bladder is emptied before

lying down and the specimen is collected on arising so that the urine collected only

reflects the

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recumbent position. Any urine voided during the night should be collected and pooled

with the first morning voided specimen.

3. Fasting specimen

This differs from a first morning specimen by being the second voided specimen

after a period of fasting.

4. 2-Hour postprandial specimen

The patient should be instructed to void shortly before consuming a routine meal

and to collect a specimen 2 hours after eating.

5. 24-hour (or timed) specimen

To obtain an accurately timed specimen, it is necessary to begin and end the

collection period with an empty bladder. The following instructions for collecting a 24-

hour specimen can be applied to any timed collection (consult test requirements to

determine if a special preservative is required):

● Day 1 – 7 AM: Patient voids and discards specimens. Patient collects all urine

for the next 24 hours.

● Day 2 – 7 AM: Patient voids and adds this urine to the previously collected Urine.

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6. Catheterized specimen

This specimen is collected under sterile conditions by passing a hollow tube

through the urethra into the bladder.

7. Midstream “clean catch” specimen

This specimen provides a safer, less traumatic method for obtaining urine for

bacterial culture. It also offers a more representative and less contaminated specimen

for microscopic analysis than the random specimen. Adequate cleansing materials and a

sterile container must be provided for the patient. The procedure for the collection of a

“clean catch” urine is described below in section VI of this policy.

8. Suprapubic aspiration

Urine may be collected by external introduction of a needle into the bladder. It is

free of extraneous contamination and may be used for cytologic examination.

9. Pediatric specimens

This may be a sterile specimen obtained by catheterization or by suprapubic

aspiration. The random specimen may be collected by attaching a soft, clear plastic bag

with adhesive to the general area of both boys and girls.

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COLLECTION PROCEDURE

I. Patient preparation:

For FEMALE patients:

1. Wash hands thoroughly before beginning the procedure and put on disposable

gloves.

2. Use betadine swabs or Hibiclens to cleanse the perineal area.

a. Separate the folds of the labia and wipe the betadine swab or Hibiclens

from front to back (anterior to posterior) on one side, then discard swab

or towelette.

b. Using a second betadine swab or Hibiclens, wipe the other side from front

to back, then discard.

c. Using a third betadine swab or Hibiclens, wipe down the middle from front

to back, then discard.

d. Pat dry periurethral area with clean dry gauze to remove excessive

betadine while keeping the labia separated.

For MALE patients:

1. Wash hands thoroughly before beginning the procedure and put on disposable

gloves.

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 2. If the patient is not circumcised, pull the foreskin back (retract the foreskin) on

the penis to clean and hold it back during urination.

3. Using a circular motion, clean the head of the penis with betadine swabs or

Hibiclens. Discard the swab or towelette.

II. Urination should begin, passing the first portion into the bedpan, urinal, or toilet.

III. After the flow of urine has started, the urine specimen container should be placed

under the patient collecting the midportion (midstream “clean catch”) without

contaminating the container.

IV. Any excess urine can pass into the bedpan, urinal, or toilet. E. Cover the urine

container immediately with the lid being careful not to touch the inside of the container

or the inside of the lid.

V. Transfer urine to specimen tube if tubes are used for transport instead of urine

containers.

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VI. Attach label to tube or container and place specimen in the transport bag. H.

Remove gloves and wash hands.

VII. Record date and time of collection and initials of the person collecting (or

submitting) the specimen on the specimen container. Transport specimen to the

Laboratory within 2 hours of collection or refrigerate and transport to the lab as soon as

possible.

SPECIMENS FOR PREGNANCY TESTING

First morning specimens are the best for pregnancy testing because the urine is

more concentrated.

VIII. LIMITATIONS OF PROCEDURE

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 A. Specimens submitted in syringes will not be accepted.

B. Specimens improperly labeled must be discarded and recollected.

C. Urine osmolality cannot be collected with preservatives.

D. Urine samples leaking in the collection bag are unacceptable.

Packaging a urine container

1. When the urine is collected the samples will be then labeled with the client’s full

name, date of birth, and the patient’s number. Write the time and date of the

collection and then stick the label on the container.

2. Note that the sample won’t be tested if the client’s name is not identified and not

labeled.

3. The nurse will then put the container in a resealable plastic bag.

4. Within the 30 minutes after the collection the sample must be placed in the

laboratory.

How to label the specimen:

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Labeling the container before taking the sample makes the packaging more easy.

A. The client’s complete first and last names, correctly spelled.

B. Write at least one of the following unique identifiers

1. Date of birth

2. Client’s ID

3. Medical record

C. Guide for proper labeling

1. Date of sample collection (MM/DD/YYYY) – write this AFTER the

collection

2. Time of sample collection (include a.m. or p.m.) – write this AFTER the

collection

Figure 1. Urine sample cup. Figure 2.

universal containers https://www.google.com/amp/s/identifydiagnostics.c.

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 https://www.kisker-biotech.com/frontoffice/product?produitId=0H-10-03

Packaging solutions

1. Peecanter urine sample collection cup - A discreet and handy pop-up urine

collection cup. It is supplied flat-packed, which reduces postal costs and storage

space, and makes it ideal for inclusion into home testing kits. The PeeCanter is

simple and quick to assemble, simply press with finger and thumb in the marked

areas and the 3D cup locks into position. The wide aperture makes it easier to

collect the sample, reducing spillages, and the square design of the cup provides

a natural and accurate pouring spout for decanting into the final urine storage or

transport container. The design eliminates the risk of crushing or cracking during

the delivery and collection process, thus increasing the potential for return of a

usable sample. The PeeCanter collection device is manufactured from card lined

with a waterproof plastic sleeve. The collection cup measures 50 x 50 x 120mm

(w x d x h) in its fully opened 3D shape, and the maximum fill capacity is 200ml.

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 2. 30 ml universal containers - Alpha Laboratories 30ml universal containers are

aseptically manufactured from clear polystyrene. All have a conical base and are

self-standing. This is a white cap with 30 ml capacity, very suitable for urine

samples and for sterile aspurates but not for sputum samples.

3. Packaging for universal tubes - Universal tubes are commonly used for urine

samples. You can transport 1 or 3 tubes in these specifically designed SpeciSafe

packs.

4. Secondary packaging for air road or road transport - Easy-to-use and economical

solutions to ensure UN3373 secondary sample packaging compliance, including

the innovative SpeciSafe® all in one system.

Urine Specimen Transport

In order to ensure proper stability of the specimen these guidelines should be followed:

I. Ensure the transport container lid is secure and leak resistant (This is

especially important, if the specimen will be sent to the laboratory through a

pneumatic tube system).

II. Utilize urine containers that are made of break-resistant plastic instead of

glass.

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 III. Verify that the specimen has been properly labeled, using at least two forms

of positive patient identification (eg, full name and hospital or medical record

number or for outpatients, full name and date of birth).

IV. Verify that the time the specimen was collected is documented (urinalysis

specimens must be analyzed within 2 hours of collection).

Collection and Transport Guidelines

I. All urine collection and/or transport containers should be clean and free of

particles or interfering substances.

II. The collection and/or transport container should have a secure lid and be

leak-proof. Leak-proof containers reduce specimen loss and risk of

healthcare worker exposure to the specimen while also protecting the

specimen from contaminants.

III. The use of containers that are made from break-resistant plastic is

strongly recommended.

IV. The container material should not leach interfering substances into the

specimen.

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 V. Specimen containers must not be re-used.

VI. Specimen tubes should be compatible with automated systems and

instruments used by the laboratory.

VII. Collection containers and/or specimen tubes should be compatible with

pneumatic tube systems where these are used for urine specimen

transport. Use of leak-proof containers is essential in this situation.

VIII. Primary (routine) specimen containers to have a wide base and a capacity

of at least 50 mL.

IX. 24 hour specimen containers to have a capacity of at least 3 litres.

X. Sterile collection containers for all microbiology specimens

XI. Specimen containers to have secure closures to prevent specimen loss

and to protect the specimen from contaminants.

XII. Amber colored containers for specimens required for assay of light

sensitive analytes such as urobilinogen and porphyrins.

Figure 3. Urine sample figure 4. Frozen urine sample

Ploy/mphotostock.com

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https://greenfleets.org/blog/store-urine-drug-test/

Storage and preservation guidelines for Urine samples

a. If the specimen cannot be processed within 2 hours of collection, for microbial

urine testing it is recommended to use chemical preservatives. Otherwise, these

specimens should be refrigerated at 2-8 degrees celsius.

b. For urinalysis it is recommended for the evaluation of urine preservation systems

by the laboratory before being utilized in the facility.

c. The proper specimen-to-additive ratio must be maintained when using a

chemical preservative to ensure accurate test results.

d. Maintain the correct ratio because it is important when transferring samples into

a preservative tube.

e. The indicated fill lines on the tube are used to ensure proper fill. Underfilling the

tube will leave a high concentration of preservative in the specimen, while

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 overfilling the tube it will overly dilute the preservative. In either case, the

function of the preservative may be compromised.

f. Evacuated tube system is designed to achieve proper fill volume to ensure the

proper specimen-to-additive ratio and proper preservation function. This also

reduces the potential exposure of the healthcare worker to the specimen.

g. It is important to note that chemical preservatives should be non-mercuric and

environmentally friendly.

1.2 SPUTUM

Sputum is the thick mucus or phlegm that is expelled from the lower respiratory

tract (bronchi and lungs) through coughing; it is not saliva or spit. Care must be taken

in the sample collection process to ensure that the sample is from the lower airways

and not from the upper respiratory tract.

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DSSM/AFB and geneXpert/ MTB:

A.Specimen Collection:

A. The number of sputum specimens to be submitted depends on the reason for

examination and the test requested.

1. Two specimens, submitted within three working days, are required for diagnosis

by DSSM, and baseline testing by TB Culture and DST. If only one specimen was

submitted for diagnosis by DSSM, and the result turned out “0” (zero), it will be

reported as “Incomplete”, and a new set of specimens will have to be submitted.

The specimens can be collected either by:

i. Spot-Early Morning Collection – the first specimen (spot) is collected when

the presumptive TB presents to the clinic. He/she is then given a sputum

container for the second specimen that should be collected early morning the

next day, and submitted to the clinic promptly.

ii. Spot-Spot (Frontloading) Collection – these specimens are collected

within the same day, at least one hour apart.

2. Only one specimen, preferably an early morning one, is needed for follow-up

examinations, Xpert MTB/RIF Assay, and LPA.

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 Note: For hospital inpatients, it is better to collect a sputum specimen each

morning on two consecutive days.

B. Sputum collection may pose a risk and should be performed with extra precaution.

Thus, collection should only be done in any of the following areas:

1. Open space (e.g. outside the DOTS facility, away from people and traffic)

2. Well-ventilated area with patient facing away from the wind.

3. Sputum collection booth – a specialized cubicle equipped with negative

pressure, HEPA filters and UV light used to prevent the spread of TB during

collection.

Note: Never collect sputum specimens inside the laboratory or in closed spaces like

toilet cubicles, waiting rooms, reception rooms, and any other poorly ventilated area.

No one should be standing in front of the patient during expectoration.

C. At least one sputum specimen should be collected under the supervision of a health

worker to guarantee correct identity of specimen and to ensure that the patient closely

follows the proper collection procedure.

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Figure 1. Ideal Sputum Container

https://images.app.goo.gl/4GzzjhoyKxK1H8hw5

D. The ideal sputum container to be used should possess the following characteristics:

1. Volume capacity of 50 mL

2. Made of transparent or translucent material

3. Wide-mouthed (at least 35mm in diameter)

4. Screw-capped

5. Unbreakable and leak-proof

6. Clean and sterile (preferably)

7. Single-use, combustible material

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 8. With walls that can be easily labeled

E. Labeling of sputum container should be done after collection. Labels should be

placed on the container’s body, not on the lid to avoid specimen mismatch.

F. An early morning specimen is an optimal specimen. This is usually collected at home.

The site staff should instruct the patient on proper collection procedure, including

correct labeling of sputum container. Sputum collected at home is collected early in the

morning so that the specimen could be delivered to the laboratory within the same day

of collection.

G. Sputum production may be induced with the use of a nebulizer containing saline

solution (5-10% sodium chloride in water) in cases when patient is having difficulty in

producing sputum.

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 Figure 2. Purulent sputum Figure 3. Mucoid Sputum

Figure 4. Blood-stained sputum Figure 5. Salivary sputum

https://images.app.goo.gl/p3WMs371ZPNpt1r89

H. Good quality sputum specimens are those that are purulent (Figure 2), mucoid

(Figure 3) and blood-stained (Figure 4). However, grossly bloody or pure blood

specimens should not be examined.

I. Poor quality sputum specimens are those that are thin, watery and composed largely

of bubbles (Figure 5). When possible, the patient should be encouraged to try collecting

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again. If not, these types of specimen can still be processed, provided that the poor

quality of the samples is reported on the result forms.

J. Prior to initiating sputum collection, the following should be observed:

1. Identify the patient.

2. Check the laboratory request form. Fill in any missing details.

3. Discuss the collection procedure as well as the reason for examination with

the patient

4. If dentures are present, advise patient to remove them and rinse mouth with

water.

5. The desired specimen is produced by a deep cough, and is thick, mucoid,

white-yellow and sometimes blood-tinged. It comes from the lower airways of

the lungs.

6. Saliva or nasal secretions are not sputum, and therefore unsuitable specimens.

7. An adequate sputum specimen with optimal quality is important to ensure

accurate and reliable test results.

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 8. Demonstrate how to properly open and securely close the specimen container

to avoid contamination. Instruct patient not to touch the inside of the container

or its lid.

K. During collection, the following should be observed:

1. If able, instruct the patient to stand. Give the patient a glass of water to rinse

the mouth free of food particles. Instruct the patient to rinse twice.

2. Instruct the patient to produce sputum by three repeated deep inhalation and

exhalation followed by a forceful cough to produce 3-5mL of sputum for each

sample.

3. If patient is unable to cough spontaneously, instruct the patient to take

several deep breaths and hold breath momentarily. Repeating this several times

may induce coughing.

4. Place the open container close to the mouth to collect the sputum

L. After collection, close the container with the screw-on lid without touching the inside

of the lid. Avoid spills or soiling the outside of the container.

M. Check the quality and quantity of the sputum produced. When possible, repeat the

procedure until desired specimen consistency and volume are met.

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N. Label the body of the sputum container with the patient’s name, date & time of

collection, and specimen number.

Note: If a patient is unable to cough spontaneously, instruct the patient to take several

deep breaths and hold breath momentarily. Repeating this several times may induce

coughing.

B. Specimen Packaging

TRIPLE PACKAGING SYSTEM

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http://www.ntp.doh.gov.ph/downloads/publications/guidelines/NTRL_MCSTSTT_2nd_Ed

ition.pdf

Triple packaging is required for the transport of pulmonary and extrapulmonary

specimens, as well as isolates, for TB laboratory testing. This employs the use of a

primary, secondary and tertiary receptacle.

FOR FRESH PULMONARY & EXTRA-PULMONARY SPECIMENS

A. Primary Receptacle

1. This refers to a durable, leak-proof, and sterile specimen container.

2. This must be labeled on the body, not on the lid, with patient’s name, date and time

of collection, and order of specimen. These must match with what is written on the

laboratory request form.

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3. Make sure that the lid is tightly closed, and sealed with parafilm.

4. An absorbent material should be placed around the specimen container.

5. When transporting multiple specimen containers, each of them should be wrapped

individually in a zip lock/vinyl/plastic bag.

6. Specimen container should be placed in an upright position to avoid leakage or

breakage.

7. All specimen containers should be placed inside a secondary receptacle.

B. Secondary Receptacle

1. This refers to a durable, leak-proof, watertight, properly sealed container.

2. When transporting fragile containers, a cushioning material should be placed

between the primary and secondary receptacle.

3. Avoid over-packing or placing more specimen containers than what the secondary

receptacle can accommodate.

4. In the event that no secondary container is available, the ziplock/vinyl/plastic bag

wrapping the primary receptacle may serve as secondary receptacle.

C. Tertiary Receptacle

1. This refers to a durable, and properly sealed transport box.

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2. All specimens, except CSF, require cold transport. Thus, ice packs should be placed

inside the tertiary receptacle.

3. Cushioning materials should also be placed to keep the specimens in upright position.

4. Outer walls should contain “This Way Up” handling label, and “UN 3373: Biological

Substance, Category B” hazard label.

5. The complete name and address of the sender and receiver should be properly

placed on the outer packaging, and on the airway bill.

6. Write “Biological Substance, Category B” on the airway bill.

Figure 6. “This way up” handling label Figure 7. UN 3733 label

https://images.app.goo.gl/QNQQrvM9FnFzoZsj9

https://images.app.goo.gl/SroxgPrXgFCJ3oXY6

FOR ISOLATES

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A. Primary Receptacle

1. This refers to a durable, leak-proof, sterile and screw-capped isolate tube.

2. Isolate tubes should contain patient identifier that matches what is written on the

laboratory request form.

3. The cap of the isolate tube should be sealed with parafilm.

4. An absorbent material should be placed around the isolate tube.

5. When transporting multiple isolate tubes, each tube should be wrapped individually

in a zip lock/vinyl/plastic bag.

6. Isolate tubes should be placed in an upright position to avoid leakage or breakage.

7. All isolate tubes should be placed inside a secondary receptacle.

B. Secondary Receptacle

1. This refers to a durable, leak-proof, watertight, properly sealed container.

2. When transporting fragile containers, a cushioning material should be placed

between the primary and secondary receptacles.

3. Avoid over-packing or placing more isolate tubes than what the secondary receptacle

can accommodate.

C. Tertiary Receptacle

1. This refers to a durable, and properly sealed transport box.

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2. Cushioning material should be placed inside the tertiary receptacle to keep the

specimens in upright position.

3. The transport box should contain “This Way Up” handling label, and “UN 2814:

Infectious Substance, Affecting Humans” hazard label

4. The complete name and address of the sender and the receiver should be properly

placed on the outer packaging, and on the airway bill.

5. Write “Isolates” on the airway bill.

Figure 8. UN 2814 label

https://images.app.goo.gl/SrDLZjP8mGz3s3a86

C. Specimen Transport

A. Ensure that a completely filled-out laboratory request form is available and

corresponds with each specimen.

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B. Transport specimens according to the triple packaging system

C. Sputum specimens collected in the morning should be delivered to the laboratory the

same day they were collected (within 2 hours) as much as possible. If this is not

possible (e.g. due to late afternoon collection, or long distance between sending and

receiving facilities),transport the specimens in coolers with ice packs sufficient to

contain the temperature similar to refrigeration.

D. Specimen Storage

When examinations are not performed on the site of collection:

1. Check the container if it is tightly capped and properly labeled. Make sure the

specimen identification matches with the laboratory request form.

2. Store in a cool (2-10°C), dry place away from sunlight until ready for transport to the

laboratory to avoid liquefaction. Refrigeration also reduces the growth of contaminants

in the specimen.

If refrigeration is not possible, sputum can be contained in coolers with ice packs. Do

not freeze.

3. Do not use any chemical preservative for storage.

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1.3 Vaginal Fluid

Collection of vaginal fluid

There are different methods of collecting vaginal fluid specimen:

1. Vaginal swab

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- A high vaginal swab is a medical procedure performed in Obstetrics and

Gynaecology, to commonly test for the presence of vaginal thrush, bacterial vaginosis

and trichomonas vaginalis. Vaginal swabs are taken to test for infections. A person may

have a vaginal swab if they have abnormal vaginal discharge, vaginal or pelvic pain, or

irregular bleeding. They may also have one as part of a sexually transmitted infection

(STI) check-up.

2. Cervicovaginal lavage (CVL),

- Cervicovaginal lavage (CVL), cervical and endocervical swabs have been used as

the primary sampling methods for HIV-1 detection and quantitation in clinical trials.

3. Pap test/ Pap Smear

- A Pap smear, also called a Pap test, is a procedure to test for cervical cancer in

women. A Pap smear involves collecting cells from your cervix — the lower, narrow end

of your uterus that's at the top of your vagina.

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Storage of vaginal fluid

Samples should be transported to the laboratory as soon as possible. If specimen

transport will be delayed, e.g. from primary care, specimens should be stored in a

refrigerator until transported to the laboratory.

Swabs should be sent in liquid transwab (purple top) medium for testing and should be

transported and examined as soon as possible.

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Vaginal Fluid Specimen Packaging

1. Take the aspirator out of the box. Do not throw away the box (the aspirator may

be returned to the package after sample collection).

2. Pull the plunger away from the aspirator's tip and then return it to the tip. To

remove the plunger and keep it from sticking, repeat this process a few times.

3. Push the plunger all the way down to the tip when you're done.

4. If the sample is to be delivered to a laboratory to be dispensed into a cryovial,

slide the aspirator back into the envelope, tip first, without reaching the envelope's

outside.

5. When putting the aspirator back into the envelope, make sure the tip does not

come into contact with something. The plunger should be extended at all times.

Vaginal Fluid Specimen Transport

1. Deliver the sample to the lab, where it will be stored at -70°C before shipment

(see Laboratory Processing Chart).

2. Within one hour, specimens should be transported to the laboratory. If this is not

possible, put the specimen on wet ice or refrigerate it at 4°C for up to 4 hours before it

can be transported.

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3. Seal the cryovial in a plastic baggie or similar to keep the exterior of the vial dry

if it will be put on wet ice.

1.4 PERICARDIAL FLUID

https://www.ucsfhealth.org/medical-tests/pericardial-fluid-culture

Pericardial fluid is an extra fluid around the space of the heart due to pericardial

effusion. This can cause strain to the heart that leads to abnormal pumping. A test is

done to get a small amount of fluid from the sac called Pericardiocentesis which is a

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method of extracting pericardial fluid that involves various steps. This is done to assist

in determining the cause of inflammation of the pericardium, the membrane that

surrounds the heart.

Pericardial Fluid Collection:

https://www.ucsfhealth.org/medical-tests/pericardial-fluid-culture

A step by step process is done meticulously by the healthcare team during the

collection of Pericardial fluid, this include the steps:

1. A cardiac monitor is placed to some patients to check for heart disturbances

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 2. Antibacterial soap is used to disinfect the chest tissue in order for the skin to be

prepped

3. Fluid is collected from the pericardial sac by inserting a needle into the chest and

in between the fifth and sixth intercostal space.

4. Once the needle is in place, put the syringe in place and aspirate to remove the

fluid build up in the sac. (In some cases, doctors insert the catheter after placing

the needle to drain excess fluid - this prevents the fluid to build up again)

Packaging of Pericardial Fluid:

After collecting, (1) the fluid will then be placed in a specimen collection tube. (2) The

collected sample of the fluid is later packaged on the petri dish with the use of a sterile

cotton swab. (3) Gently rub the swab over the gelled agar on the petri dish by rolling

the swab with your fingers. Lastly, (4) cover the dish with a lid and label it with the

date and the item tested.

aq2

Transport of Pericardial Fluid:

41 | P a g e
In the transportation of the specimen, the procedure are as follows:

1. Maintain transport temperature at 18-28 °C.

2. Transport to the laboratory as soon as possible.

https://direct.dksh.com.au/product-

dialog.asp?id=12403&cid=1223&linkpath=%2FProduct%2FLaboratory%2FEquipment%

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ansport%2Dcontainer%2Dclearblue%2D1%2Deach

How to store Pericardial Fluid:

1. If transport will be delayed, keep refrigerated. DO NOT FREEZE.

2. Store in sterile bottles with tight cap

3. 16 hours at room temperature (18-28°C)

4. Use a plain sterile tube or container for microbiology testing

42 | P a g e
https://www.researchgate.net/figure/Bottle-showing-the-pericardial-fluid-which-

is-yellowish-coloured-and-remained-clotted_fig3_51675055

1.5 Synovial Fluid

Synovial Fluid Collection

Synovial fluid (Arthrocentesis)

Arthrocentesis is the process by which synovial fluid collection occurs by

penetrating the joint space through aspiration. This procedure should be done

43 | P a g e
under sterile procedural conditions and performed by a physician with intimate

knowledge of the involved anatomy.

https://healthjade.net/arthrocentesis/

Indications

Synovial fluid aspiration is indicated for the following:

● Evaluation for an intra-articular infectious process

● Diagnosis of inflammatory disease (e.g., crystalline arthropathy,

spondyloarthropathies)

● Administration of medications for acute or chronic arthritis

44 | P a g e
 ● Symptom relief in a swollen, painful joint or inflammatory

conditions (e.g., rheumatoid arthritis)

● Evacuation of possible hemarthrosis in a traumatic effusion

● Identifying communication between the joint space and a laceration

Equipment

Skin Preparation

● Skin preparation solutions (e.g., chlorhexidine, betadine)

● Sterile gloves

● Sterile drapes

● Sterile gauze

Syringes

● 3 cc to 5 cc syringe for instilling local anesthetic

● 10 cc to 20 cc syringe for fluid aspiration

45 | P a g e
 Needles

● 25 gauge to 27 gauge for local anesthetic introduction

● 18 gauge to 22 gauge for fluid aspiration

Medications

● Local anesthetic

● Medications being injected into the joint, if applicable

Other

● Three-way stopcock for draining large amounts of fluid

● Specimen collection tubes for laboratory analysis

● Hemostat (for stabilizing the needle when exchanging

syringes if more than one is required for complete aspiration)

Technique

1. Define the joint anatomy by palpating the surrounding bony

landmarks. Ultrasound may be helpful in locating effusions.

46 | P a g e
2. Select a puncture site and an approach to the joint based on the

appropriate anatomy. Be sure to avoid tendons, major blood

vessels, and major nerves.

3. Apply the antiseptic solution to the area of needle insertion and

surrounding skin. Allow the skin to dry and then put down the

sterile drape surrounding the area of needle insertion.

4. Using a 25 gauge to 27 gauge needle, first create a wheal of local

anesthetic at the point of insertion. After the skin is anesthetized,

infiltrate the skin down to the area of the joint capsule. For

extremely painful joints, a regional nerve block can be used.

5. Attach a larger needle of appropriate length to an appropriately

sized syringe. Insert the needle into the joint space along the

anesthetized track.

6. For draining larger effusions, a three-way stopcock can be placed

between the needle and the syringe.

7. To change the syringe during the procedure, grasp the hub of the

needle with a sterile hemostat and hold it tightly while removing

the syringe.

8. An attempt should be made to remove as much fluid or blood as

possible. If fluid stops flowing, the joint is either drained completely,

47 | P a g e
 the needle tip is dislodged, or debris is obstructing the needle.

Slightly advance or retract the tip, rotate the bevel, or aspirate less

forcefully.

9. Place fluid into appropriate tubes and send the synovial fluid for

studies as indicated by the clinical scenario.

10. Place a dressing or bandage over the puncture site, and

apply pressure to achieve hemostasis.

Fluid Analysis

● Common laboratory analyses include cell count, gram stain, crystal

analysis, and glucose and protein levels.

● Normal synovial fluid will have the following characteristics: Clear or

straw-colored, less than 200 leukocytes per microL, variable neutrophil

count, and negative gram stain.

48 | P a g e
 ● Inflammatory synovial fluid will have the following characteristics: Yellow

color, 2,000 to 50,000 leukocytes per microL, variable neutrophil count,

and negative gram stain.

● Hemorrhagic synovial fluid will have the following characteristics: Opaque

appearance, less than 2,000 leukocytes per microL, less than 25%

neutrophils, negative gram stain.

● Septic synovial fluid will have the following characteristics: Cloudy/opaque

appearance, greater than 50,000 leukocytes per microL, greater than 90%

neutrophils, and a positive gram stain between 30% to 80% of the time.

Synovial Fluid Packaging

Each specimen must be accompanied by a completed and matching Synovial

fluid cytology request form.

All specimen containers must be clearly labelled with:

1. Patient’s full name

2. Date of birth

3. Hospital number

49 | P a g e
Requires three layers of packaging:

● Primary container (e.g. universal tube, vial)

● Secondary container (e.g. specimen bag)

● Outer packaging (e.g. rigid transport box).

The primary sample must be individually bagged in a secondary bag and sealed.

If the sample is liquid, enough absorbent material must be added to the

secondary bag to absorb a potential spillage of the sample. The request form

must be placed in the specimen bag’s separate pouch. Specimens must then be

placed in a rigid box and closed. The box must comply with Transport

Regulations.

Synovial Fluid Transportation

The fluid should then be transferred to a sterile specimen collection cup and

sealed for transport to the laboratory for analysis. The syringe may be emptied

50 | P a g e
and reattached to the indwelling needle repeatedly, minimizing needle entry

points. Once collected into a suitable anticoagulant tube, the specimen must be

received in the laboratory in the shortest possible time. Synovial fluid samples

requiring transport on the public road must be packaged and transported in

compliance with “The Carriage of Dangerous Goods and Use of Transportable

Pressure Equipment Regulations (ADR Regulations) 2011.” Specimens must be

delivered to the laboratory within 24 hours. If there is unavoidable delay in

sending the specimen, it should be refrigerated at 4°C.

Storage of Synovial Fluid Specimens

Although synovial fluid can be stored overnight in a 4°C refrigerator without

significantly degrading in cytological appearances, it is best to make cytological

preparations as soon as possible after aspiration5. Under no conditions should

the fluid be frozen.

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1.6 PERITONEAL FLUID

Peritoneal fluid is a liquid that acts as a lubricant in the abdominal cavity. It is

found in small quantities (generally 5-20 mL) between the layers of the peritoneum that

line the abdominal wall. Peritoneal fluid acts to moisten the outside of the organs and to

reduce the friction of organ movement during digestion and movement. To help

diagnose the cause of peritonitis, an inflammation of the membrane lining the abdomen,

and/or peritoneal fluid accumulation, where fluid builds up in the abdomen or around

internal organs.

52 | P a g e
 https://ssl.adam.com/graphics/images/en/9737.jpg

Collection of Specimen for Gram Stain and Culture Sensitivity (Paracentesis)

Abdominal tap (paracentesis) is used to remove fluid from the area between the

belly wall and the spine. This space is called the abdominal cavity or peritoneal cavity. It

has 2 kinds namely:

● Diagnostic tap- small amount of fluid is taken and sent to the

laboratory testing.

● Large volume tap- Several liters may be removed to relieve

abdominal pain and fluid build-up.

53 | P a g e
1. Empty your bladder either voluntarily before the Abdominal Tap Procedure or

with a Foley Catheter. Make sure the patient is in the horizontal supine

position, and tilt the patient slightly to the side for collection. Take note of the

insertion sites.

https://5minuteconsult.com/collectioncontent/30-156350/procedures/abdominal-

paracentesis

2. Prepare the skin with povidone-iodine or chlorhexidine solution and allow it to

dry while applying sterile gloves and a mask.

3. Center the sterile drape about one third of the distance from the umbilicus to

the anterior iliac crest.

54 | P a g e
https://5minuteconsult.com/collectioncontent/30-156350/procedures/abdominal-

paracentesis

4. Infiltrate the skin and subcutaneous tissue with a 1% solution of lidocaine

with epinephrine. A 2-inch needle is then inserted perpendicular to the skin to

infiltrate the deeper tissues and peritoneum with anesthetic.

5. Insert the catheter/introducer through the skin. The nondominant hand then

stretches the skin to one side of the puncture site, and the needle is further

inserted to create a Z tract.

https://5minuteconsult.com/collectioncontent/30-156350/procedures/abdominal-

paracentesis

6. Advance the catheter until a “pop” is felt and the catheter penetrates the

55 | P a g e
 peritoneum. Release the pressure on the skin after the introducer enters the

peritoneum. Advance the catheter into the abdominal cavity.

7. Remove the introducer, and attach the syringe. Draw the fluid into the

syringe. If no fluid returns, rotate, slightly withdraw, or advance the catheter

until fluid is obtained. If still no fluid returns, abort the procedure, and try an

alternative site or method.

· Alternative method: Attaching a three-way stopcock or one-way

valve. If lavage is desired, connect intravenous tubing to the

three-way stopcock.

https://5minuteconsult.com/collectioncontent/30-

156350/procedures/abdominal-paracentesis

8. After the procedure, gently remove the catheter, and apply direct pressure to

the wound. If the insertion site is still leaking fluid after 5 minutes of direct

pressure, suture the site with a vertical mattress suture and perform aftercare.

9. Observe the characteristics of the fluid and transport it to the laboratory for
56 | P a g e
 gram staining to see a stain in order to determine and classify into 2 bacterial

species (gram-positive bacteria) and (gram-negative bacteria) and culture

sensitivity to see how bacteria grows and to get organisms that may cause of

the infection.

PACKAGING:

1. Inoculate a minimum of 3 mL into an Aerobic and Anaerobic Blood Culture Bottle.

2. The patient's full name, date of birth or health card number, the source of the

fluid and date and time of collection should be specified on the requisition and

specimen container.

TRANSPORT

Specimen processing instructions: Obtain peritoneal fluid (minimum 50 mL) in

57 | P a g e
 specimen container.

Transport temperature: Refrigerate or keep on wet ice until transported to

laboratory.

STORAGE

If transport will be delayed, keep refrigerated but do not freeze it. For

specimen stability and storage, store it for 16 hours at room temperature between 18 -

28 °C.

1. 7 SEMINAL FLUID

A complete semen analysis measures the quantity and quality of the fluid

released during ejaculation. It evaluates both the liquid portion, called semen or

seminal fluid, and the microscopic, moving cells called sperm. It is often used in the

evaluation of male infertility.

COLLECTION

1. Refrain from any sexual activity (including masturbation) for at least 2 days

58 | P a g e
 and no more than 10 days. Longer or shorter periods of abstinence may

result in a lower sperm count or decreased sperm motility.

2. A private room is available for semen collection within close proximity of our

laboratory and is recommended for collection.

3. The semen specimen should be obtained by masturbation. Alternate methods

may be discussed but are not recommended. But, if you don’t have sperm or

have a severe male infertility, it may require a surgical procedure such as

microsurgical epididymal sperm aspiration (MESA) or testicular sperm

aspiration (TESA). Do not use any lubricant, including saliva, when collecting

semen. Do not collect the specimen in a condom as these contain spermicidal

agents, which will alter the results of the analysis. If a condom must be used,

the laboratory will provide a special semen collection condom.

4. The specimen should be collected in a container. Be sure hands and penis are

cleaned prior to collection. Avoid touching the inside of the cup. If any semen

is spilled, DO NOT attempt to transfer it to the cup. Inform the lab personnel

about the spill.

59 | P a g e
5. If the specimen was obtained outside of the collection room, bring the

specimen to the laboratory within one hour after ejaculation. Do not expose

the specimen to extremes of temperature. Place the specimen container

upright in a plastic bag, with the lid securely tightened, and keep specimen

close to body temperature by transporting close to the body. The specimen

should not be placed in a purse, pocket, or briefcase. Sperm do not have a

long life outside of the body and at different temperatures. Delays in

delivering semen and exposure to various temperatures will result in lower

overall motile sperm count and poor semen cryopreservation.

6. Semen older than 1 hour will not be accepted for cryopreservation.

7. Please label the specimen collection container with the self-adhesive label

printed with your name, birthdate and date and time of collection.

8. SPECIMENS THAT ARE NOT ACCEPTABLE:

a. the container has a crack, is missing, or is leaking.

b. container not licensed by the ART lab

c. container is hot or cold to the touch

60 | P a g e
 d. specimen obtained with an unapproved condom

e. specimen brought in by unscheduled walk-in appointments

https://www.freepik.com/premium-photo/container-biomaterials-with-sperm-

analysis_10255998.htm

PACKAGING

1. Place the specimen container upright in a plastic bag with the lid tightly

fastened, and transport the specimen close to the body to hold it at body

temperature.

2. It is not recommended that the specimen be carried in a purse, bag, or

briefcase.

3. Outside of the body and at various temperatures, sperm do not have a long

life.

61 | P a g e
 4. Lower overall motile sperm count and poor semen cryopreservation will result

from delays in delivering semen and exposure to various temperatures.

http://what-when-how.com/nursing/specimen-collection-client-care-nursing-part-1/

TRANSPORT

Specimens must be packed for transport in a way that prevents cross

contamination of forms or other specimens when a specimens leak. Specimens must be

transported in such a way to ensure the safety of the courier, the general public and

the receiving laboratory:

1. During transport, do not agitate the container in any way.

62 | P a g e
 2. During transport to the laboratory, the sample should be kept between 20 °C

and 37 °C. Avoid direct sunlight, extreme heat or cold.

3. The sample can be kept warm by keeping the container close to the body. Sperm

do not have a long life outside the body and at different temperatures.

4. If the specimen was obtained outside of the collection room, the report should

note that the sample was collected at home or another location outside the

laboratory. Then, immediately bring the specimen to the Outpatient Laboratory

one hour after ejaculation for billing and paperwork processing.

5. Delays in delivering semen and exposure to various temperatures will result in

lower overall motile sperm count and poor semen cryopreservation.

Storage

Semen should be stored at a constant temperature of 17°C - 2°C to maintain

semen viability and maximize shelf life. Semen is extremely temperature sensitive:

shelf life is shortened at temperatures above 20°C; while temperatures below 15°C

are likely to reduce sperm viability. Semen doses should always be treated carefully

to prevent damage from handling and protected from exposure to light.

63 | P a g e
 1.8 CEREBROSPINAL FLUID

Description

Cerebrospinal fluid is defined as a clear, colorless body fluid that surrounds the brain

and spinal cord of all vertebrates. It is produced by specialized ependymal cells. It

serves to act as a cushion or a buffer, at the same time, providing protection and

64 | P a g e
support to the brain inside the skull.

Figure 1 CSF System

Biosafety

It is important to adhere to proper biosafety guidelines while handling potentially

infectious clinically specimens in order to maintain a safe working environment for

patients, health care workers, and laboratorians. Infection may be transmitted from

patient to staff and from staff to patient during the procedures described. In addition to

the agents that cause bacterial meningitis, the patient could have other bacterial or

viral agents in either the CSF of blood and both are great hazards and potentially lethal.

Of particular importance are the viruses causing hepatitis and acquired

65 | P a g e
immunodeficiency syndrome. To decrease the risk of transmission of these agents, the

recommendations below should be followed:

● Wear latex or nitrile gloves that are impermeable to liquids and change gloves

between every patient

● Dispose of syringes and needles in a puncture resistance, autoclavable discard

container. Do not attempt to re-cap,shear, or manipulate any needs. A new

sterile syringe and needle must be used for each patient.

● For transport to a microbiology laboratory, place the specimen in a container that

can be securely sealed. Wipe any bottles with CSF or blood on the outside

thoroughly with a disinfectant, such as a 70% alcohol swab.

○ Do not use povidone-iodine on the rubber septum of a T-I or blood culture

bottle

● Remove gloves and discard in an autoclavable container

● Wash hands with antibacterial soap and water immediately after removing gloves

● In the event of a needle-stick injury or other skin puncture or wound, wash the

wound liberally with soap and water. Encourage bleeding.

● Report a needle-stick injury, any other skin puncture, or any contamination of

the hands or body with CSF to the supervisor and appropriate health officials

66 | P a g e
 immediately as prophylactic treatment of the personnel performing the

procedure may be indicated

II. Collection and transporting of CSF

The collection of CSF is an invasive procedure and should be only performed by

experienced personnel under aseptic conditions. If bacterial meningitis is suspected,

CSF is the best clinical specimen to use for isolation, identification, and characterization

of the etiological agents. Suspected agents should include N. meningitidis, S

pneumoniae, and H influenza and other pathogens in some cases.

a. Preparing for lumbar puncture

If possible, three tubes (1 ml each) of CSF tube of CSF is available, it should be given to

the microbiology laboratory. Because the presence of blood can affect cultures of CSF,

if more than one tube of CSF is collected from a patient, the first tube collected (which

could contain contaminating blood from the lumbar puncture) should not be the tube

sent to the microbiology laboratory. The kit for collection of CSF should contain

● Skin disinfectant: 70% alcohol swab and povidone-iodine

67 | P a g e
 ○ Alcohol with concentrations greater than 70% should not be used because

the increased concentrations result in decreased bactericidal activity. Do

not use alcohol with glycerol added to it.

● Sterile gloves

○ Be sure to check the expiration date:

● Sterile gauze

● Adhesive bandage

● Lumbar puncture needle

○ 22 gauge/89mm for adults

○ 23 gauge/64 mm for children

● Sterile screw-cap tubes

● Syringe and needle

● Transport container

● T-I medium ( if CSF cannot be analyzed in a microbiology laboratory immediately)

○ T-I should be refrigerated at 4 degrees celsius and added to the kit

immediately before use in the field.

● Venting needle (only if T-I is being used)

● Instructions for lumbar puncture and use of T-I medium

68 | P a g e
 Figure 2 CSF Storage Materials

b. Lumbar puncture procedure

Follow all appropriate biosafety precautions

1. Gather all materials from the CSF collection kit and a puncture-resistance

autoclavable container for used needles

2. Wear surgical mask and sterile latex or nitrile gloves that are impermeable to

liquids and change gloves between every patient

3. Label the collection tubes with appropriate information: patient’s name, date and

time of specimen collection, and Unique Identification Number. Be sure this

number matches the number on both the request and report forms

69 | P a g e
4. Ensure that the patient is kept motionless during the lumbar puncture procedure ,

either sitting up or lying on the side, with his or her back arched forward so that

the head almost touches the kneed in order to separate the lumbar vertebrae

during the procedure

5. Disinfect the skin along a line drawn between the crests of the two ilia with 70%

alcohol and povidone-iodine to clean the surface and remove debris and oil.

Allow to dry completely.

6. Position the spinal needle between the 2 vertebral spines at the L4-L5 level and

introduce into the skin with the bevel of the needle facing up

a. Accurate placement of the needle is rewarded by a flow of fluid, which

normally is clear and colorless

7. Remove CSF (1 ml minimum, 3-4 ml if possible) and collect into sterile screw-cap

tubes. If 3-4 ml CSF is available, use 3 separate tubes and place approximately

1ml into each tube

8. Withdraw the needle and cover the insertion site with an adhesive bandage.

Discard the needle in a puncture-resistant, autoclavable discard container

9. Remove mask and gloves and discard in an autoclavable container

10.Wash hands with antibacterial soap and water immediately after removing gloves.

11.Transport the CSF to a microbiology laboratory within 1 hour for culture analysis.

70 | P a g e
 a. If that is not possible, inoculate CSF into T-I medium ( see Section I.C.

below).

b. If TI is not available, incubate CSF at 35-37 degrees Celsius with ~5%

CO2 and store in an approved location if the laboratory is closed.

12.In the event of a needle-stick injury or other skin punctured or wound, wash the

wound liberally with soap and water. Encourage bleeding.

13.Report a needle-stick injury, any other skin puncture, or any contamination of

the hands or body with CSF to the supervisor and appropriate health officials

immediately as prophylactic treatment of the personnel performing the

procedure may be indicated.

71 | P a g e
Figure 3 CSF Extraction

C. Inoculating and transporting T-I medium

T-I is a biphasic medium that is useful for the primary culture of meningococci and

other etiological agents of bacterial meningitis (S. pneumoniae and H. influenzae) from

CSF. It can be used as a growth medium as well as a holding and transporting medium.

The preparation of T-I media is described in the Annex. T-I media should be stored at 4

degrees Celsius and warmed to room temperature (25 degrees Celsius) before use.

1. Label the T-I bottle with appropriate information: patient name, date and time of

CSF inoculation, and Unique Identification Number. Be sure this number matches

the number on both the request and report forms.

2. Use sterile forceps to pull the aluminum cover of a T-I bottle away from the

rubber stopper and disinfect the stopper with 70% alcohol. Allow to dry.

a. Do not use povidone-iodine as it may be carried into the medium by the

passing needle and would inhibit growth of bacteria.

b. Do not completely remove the aluminum cover

3. Use a sterile syringe and needle to inoculate 0.5-1.0 ml of CSF into the T-I

medium. The remaining CSF should be kept in the collection tube. It should not

be refrigerated, but should be maintained at room temperature (20-25 degrees

72 | P a g e
 Celsius) before Gram staining and other tests. Discard the needle in a puncture-

resistant, autoclavable discard container.

4. After inoculation, invert the T-I bottle several times to mix.

5. If transport to a reference laboratory is delayed (next day or longer), insert a

venting needle (sterile cotton-plugged hypodermic needle) through the rubber

stopper of the T-I bottle, which will encourage growth and survival of the

bacteria.

a. Be sure that the venting needle does not touch the broth.

6. Incubate inoculated T-I medium at 35-37°C with ~5% CO2 (or in a candle-jar)

overnight or until transport is possible. If transportation is delayed more the 4

days, remove the vented T-I bottle from the incubator or candle jar and place at

room temperature until shipment.

7. Remove the venting needle and wipe the rubber stopper with 70% alcohol

before shipping. It is essential to avoid contamination when sampling the bottles

to obtain specimens aseptically.

8. If the T-I bottle can be transported to a reference laboratory the same day, do

not vent the bottle until it arrives in the receiving laboratory. Upon arrival, vent

the T-I bottle, incubate at 35-37°C with ~5% CO2 (or in a candle-jar), and

observe daily for turbidity in the liquid phase for up to 7 days.

73 | P a g e
 a. If turbidity is observed, culture onto a blood agar plate (BAP) and a

chocolate agar plate (CAP) immediately (see Chapter 6: Primary Culture

and Presumptive ID).

b. If no turbidity is observed, culture onto a BAP and a CAP on day 4 and

day 7.

c. If T-I medium appears to be contaminated, selective media such as

Modified Thayer-Martin and chocolate agar with bacitracin may be used.

d. Transporting CSF specimens without T-I media

CSF specimens should be transported to a microbiology laboratory as soon as possible.

Specimens for culture should not be refrigerated or exposed to extreme cold, excessive

heat, or sunlight. They should be transported at temperatures between 20°C and 35°C.
74 | P a g e
For proper culture results, CSF specimens must be plated within 1 hour for instructions

on processing CSF once it has arrived in the laboratory.If a delay of several hours in

processing CSF specimens is anticipated and T-I medium is not available, incubating the

specimens (with screw-cap loosened) at 35-37°C with ~5% CO2 (or in a candle-jar)

may improve bacterial survival.

Packaging of Cerebrospinal fluid

The packing must be of good quality, strong enough to withstand the shocks and

loadings normally encountered during transport, including trans-shipment between

transport units and warehouses as well as any removal from a pallet or overpack for

subsequent manual or mechanical handling.

The packaging must consist of three components 1) a primary receptacle (the tube, or

other container typically made of glass or rigid plastic, including the stopper, cap or

other closure elements, that is in direct contact with the specimen); 2) a leak-proof

secondary packaging; and 3) a rigid outer packaging

75 | P a g e
Absorbent material must be placed between the primary receptacle and the secondary

packaging

One external surface of the outer packaging clearly must show the text “BIOLOGICAL

SUBSTANCE, CATEGORY B” Adjacent to this, inside a diamond mark, must appear the

text “UN 3373”

If shipping frozen Samples with dry ice, the outer packaging must be marked with the

text “Dry Ice'' or “Carbon dioxide, solid” and “UN 1845” and the net quantity, in

kilograms, of dry ice. These markings must be accompanied by the Class 9 label for

Miscellaneous Dangerous Goods.

Storage of Cerebrospinal Fluid

Guidelines on storage of cerebrospinal fluid indicate that collected bacterial culture

samples should be stored at temperatures ranging from 20°C to 35°C since culture

samples might not be able to survive extreme temperatures while non-culture samples

should be cooled down to about +4°C as soon as possible and then frozen at about -

20°C preferably on the same day as to when the sample was collected. If storage for

the sample is for long term, it must be frozen to about -80°C or colder (e.g. liquid

76 | P a g e
nitrogen). Sample storage containers have specific measurements being 13mm in

diameter in tube.

1.9 GASTRIC FLUID

Gastric fluid is a unique combination of hydrochloric acid, lipase, and pepsin. Its main

function is to inactivate swallowed microorganisms, thereby inhibiting infectious agents

from reaching the intestine. The mucosa in the mouth and the esophagus is squamous

epithelium which, like the skin, could be presumed to protect against infection. On the

other hand, the epithelium in the stomach and gut is of the simple columnar type,

which would be expected to be more easily penetrated by infectious agents. The gastric

fluid is thus the first line of defense against infection throughout the gastrointestinal

tract.

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https://image.shutterstock.com/image-photo/bile-yellowgreen-liquid-secreted-by-

260nw-789041833.jpg

COLLECTION

Cytology Requisition Information:

All specimens must be submitted for testing with a completed Cytology & HPV

Testing Requisition. Provide the following information in a legible format:

Patient Information:

• Full name of patient (printed in the same format as patient’s health card)

• Health card number

78 | P a g e
 • Date of birth

• Date of specimen collection

• Specimen source

• Specimen site

• Number and type of specimen (e.g. slides, vials)

• Collection method (e.g. voided urine, fine needle aspiration)

• Pertinent clinical information

Health Care Provider Information:

• Full name, address and billing number of the ordering health care

provider.

• Full name, address (and billing number if known) of any copy-to

physicians.

Collection Kit Information:

Fine Needle/ Sputum/ Fluid Kits: Kit components are ordered separately:

o 90 mL sterile urine container with 40 mL of cytology preservative

(Cytolyt ®- clear, colourless solution).

79 | P a g e
 https://catalog.hardydiagnostics.com/cp_prod/product/Cat

Nav.aspx?oid=514&navpath=1917,8174&prodoid=PC4090400S

o Collection instructions are included in the sputum and urine kits

https://ecog.io/magnifierq/index.php?keyword=30ml-Universal-Container-

With-Label-229574

80 | P a g e
o Polybag

https://www.shorr.com/who-we-serve/medical-packaging-solutions

o Cytology requisition (ordered separately)

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https://cloudpractice.freshdesk.com/support/solutions/articles/3000075561-

lifelabs-cytology-hpv-testing-requisition

CAUTION: The preservative contains methyl alcohol. Do not drink. If

ingested, do not induce vomiting; call your doctor or local poison

control center immediately. Vapor may be harmful if inhaled; use with

adequate ventilation. Flammable; keep away from heat, sparks & open

flame. Avoid contact with eyes.

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PACKAGE

Specimen Labeling:

All specimens will be clearly labeled BEFORE being sent to the laboratory for

testing to ensure correct identification of the patient and sample.

All specimens/containers must be labeled with:

• The patient’s full name (printed in the same format as patient’s health

card)

• A second identifier such as date of birth or health card number

• It is recommended that the specimen container also be labeled with

specimen source (for non-gynecologic samples)

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https://line.17qq.com/articles/uwcssqrrx.html

Specimen/container labeling options are:

• Computer printed label affixed to the side of the specimen container.

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https://www.osfhealthcare.org/lab/specimens/labeling/

• Or clearly printed handwritten information on the label of the specimen

container using indelible ink

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 https://aadpathology.com/specimen-collection-handling-guidelines/

• Specimens collected on a glass slide must have the patient/sample

information printed on the frosted end of the slide using pencil or indelible

ink.

http://www.prepared-microscopeslides.com/sale-12368534-yeast-w-m-on-

bacteria-microbiology-microspecimen-prepared-microscope-slides-for-

educational-supplies.html

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Specimen Handling:

• Specimens collected from multiple sites should be collected in separate

vials/slides with the specimen source identified.

• Each fluid specimen must be placed into a polybag.

https://www.shorr.com/who-we-serve/medical-packaging-solutions

• Specimens collected on a glass slide must be securely packaged in a

cardboard/plastic slide holder

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http://www.prepared-microscopeslides.com/sale-12368534-yeast-w-m-on-

bacteria-microbiology-microspecimen-prepared-microscope-slides-for-

educational-supplies.html

• A completed Cytology & HPV Testing Requisition must accompany each

specimen.

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https://cloudpractice.freshdesk.com/support/solutions/articles/3000075561-

lifelabs-cytology-hpv-testing-requisition

• Specimens requiring expedited service must be clearly marked as such.

The typical designation is: ASAP.

https://www.expeditedelivers.com/industries/medical-specimen-delivery/

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TRANSPORT

Specimens must be transported in a specimen cup or fluid/sputum kit placed

inside a cooler box to the laboratory for processing/storage as soon as possible.

It is noted that if necessary, it can be transported in ambient or room

temperature without comprising the sample’s stability. Standard precaution is to

be observed to prevent infection. If the specimens will be transported in more

than 4 hours, put them in the refrigerator (4–8 °C) and keep them there until

they are transported.

STORAGE

To preserve viability and improve shelf life, gastric fluid should be stored in

tightly sealed containers at 4 °C (short-term) or at −20 °C or preferably at

−80 °C (long-term). The specimen must not be exposed to the air so as to not

let microbiota to grow uncontrolled and release unrestrained vapors. The

specimen’s stability or lab retention can last up to 3 days.

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1.10 Pleural Fluid

Description

Pleural fluid is a liquid that is found in between layers of the pleura. The pleura is

defined as a two-layer membrane that covers the lungs and chest cavity. The area of

which the pleural fluid is found within the pleura is called the pleural space. For normal

findings, there is normally a small amount of pleural fluid in the pleural space.

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Collection

Figure 1: Pleural fluid extraction

Collection of Pleural fluid starts with the application of an antiseptic solution to disinfect

the location of where the extraction of the fluid is to occur. Numbing medicine is then

applied first. An empty syringe is then inserted into the lower ribs where the

anaesthesia was applied. It is then advanced until it enters the fluid collection inside the

chest wall. Depending on the amount of fluid to be extracted, a syringe is used if only a

small amount is to be extracted. For larger quantities, a tube attached to a jar is used

for the extraction. It is of utmost importance that the patient does not breathe heavily

to inhibit the lung from expanding, avoiding contact with the needle.

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TRANSPORT

Transport the specimens, depending on whether processed or unprocessed, in sterile

containers. Specimens are normally transported in ambient or room temperature. The

maximum acceptable time delay before specimens are processed in the laboratory is 2

hours. If a longer delay is expected, the specimen should be stored in a refrigerator at

4ºC, except for microbiological cultures.

STORAGE

Temperature and storage time are potential preanalytical errors in pleural fluid analyses,

particularly when glucose and LDH are unstable. The best practice is to run all of the

experiments as soon as they are collected. Except for LDH analysis, most studies can be

performed on refrigerated samples.

Due to the obvious instability of isoenzymes 4 and 5, a decrease in LDH was observed

within the first 24 hours in samples kept at -20 degrees C and after 2 days in samples

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kept at 4 degrees C. Except for glucose, all parameters remained stable at room

temperature or 4 degrees Celsius for at least four days.

PACKAGING

Preferred collection container:

They may be submitted in a dry sterile container or in a large collection device such as

a vacutainer bottle

https://www.researchgate.net/figure/Pleural-fluid-before-left-and-after-right-

centrifugation-The-centrifugation-resulted_fig1_244949069

Specimen processing instructions:

● Obtain pleural fluid (minimum 50 mL) in a specimen container.

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 ● The specimen container must be labeled. Addressograph labels are preferred,

otherwise the following information must be provided, clearly written in ink:

(a) Minimum of two Patient identifiers

(i) patient's full name

(ii) hospital unique number

(iii) date of birth

(iiii) health card number

(b) Date of specimen collection.

(c) Specimen type and site, as it is written on the Cytology Requisition form.

This information must be recorded on the side of the specimen container and not the lid.

If a specimen is known or suspected to contain unique or extreme biohazard the

container shall be so marked. The referral lab does not perform cytology testing on CJD

cases.

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156350/procedures/abdominal-paracentesis

Aldrovandi, G. M.D. (2015). Female Genital Secretions Collection Processing.

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CHONWATTANA, Famui

Lexington Medical Center. (n.d.). Semen Collection Protocol.

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Martinsen, T. C., Fossmark, R., & Waldum, H. L. (2019, November 29). The

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