Collection,Storage and Transport of Microbiological Specimen

Dr.Rubina Kamran PG trainee Microbiology Department of Pathology P.I.M.S Islamabad

GENERAL RULES FOR COLLECTION &TRANSPORTATION

Strict aseptic techniques Collect specimen before administration of antibiotics Prevent contamination from normal flora Optimum timing/ site Leak proof sterile container

GENERAL RULES FOR COLLECTION &TRANSPORTATION

Proper labeling Brief clinical history Prompt transportation to the laboratory

Chain of collection &transportation

CRITERIA FOR REJECTION OF SPECIMENS

Missing or inadequate identification Leaking container Inappropriate container Insufficient quantity Unknown time delay Inappropriate requests e.g,oral swabs,foley`s cathetar tip etc.

CRITERIA FOR REJECTION OF SPECIMENS

Incomplete forms Inappropriate transport or storage

Ways to transport specimen

Cold chain Refrigerating or by keeping cool,ice box,thermos flask. urine At 37C keep near body and put in incubators immediately or Transport media. Csf urethral,cervical ,semen blood culture

Objective of using transport Media

To keep the organisms alive suppress the growth of unwanted organisms Outgrow the no. of wanted organisms

Transport Media

Amies transport medium

Carry-Blair for stool culture

Stuart media for aerobic specimen

Conti---

Cary Blair t.medium-----Enteric pathogens. Amies t.medium------suspected anaerobic bacteria. Stuart t.media------urethral &genital specimens.

INDIVIDUAL SPECIMENS COLLECTION AND TRANSPORT

SPUTUM

A clean,dry,leak-proof container to be used.

Patient is instructed to cough deeply. Specimen must be SPUTUM not Saliva. Avoid use of mouth-wash prior to collection. Early morning samples should be collected.

Container for sputum

Transport of Sputum

Early dispatch.
Transfer purulent part of sputum to cotton-wool swab. Container of Amies transport medium. Must reach laboratory with in 6hrs.

Collection of Throat & Mouth Specimen

In good-light and using a tongue-depressor examine the inside of mouth . Look for inflammation, presence of membrane/ exudate /pus.
Swab with a sterile cotton-wool swab. Avoid contamination with saliva i.e;dont touch any-other part of mouth.

Throat specimen Collection

Avoid antiseptic gargles eight hours prior to collection. Deliver with in two hours of collection.

Alternate Procedures Trans-laryngeal aspiration/ BAL required in case of

Patient is debilitated & CANNOT EXPECTORATE spontaneously.

Routine samples have failed to recover causative organism. An-aerobic pulmonary infection is suspected. Lung-abscesses/deep-seated pulmonary infections.

Nasal swabs

Use sterile cotton wool swab,gently swab the inside surface of the nose. Take care not to contaminate the swab, replace in its sterile container Label and within 2 hrs. deliver to the lab with request form.

Ear discharges

Collect the specimen of the discharge on cotton wool swab without contaminating it. Transport within 2 hrs. to the lab. Transport it within 6 hrs if Amies transport medium is used.

Eye specimens

Should be collected by a medical officer or an experienced nurse. Cotton wool swab ,collect the discharge,& in an infant swab the lower conjunctival surface. Deliver immediately to the lab.or use Amies Transport media, also make a smear of the discharge on a slide.

Blood cultures Collection procedure

Aseptic technique like lumber puncture Take out BC bottle from fridge & bring to room temp wipe its capping with anti-septic. Identify the vein Operater hand clean . Sterile gloves Disinfect skin by standard method(Tinc iodine or Povidone iodine (1 min), 70% alcohol (not spirit). Avoid recontaminating the veinipuncture site. Use new needle to transfer blood into BC bottle.

NUMBER & TIMING

Should be drawn before use of SYSTEMIC ANTI-MICROBIAL drugs. When patient has recurring fever, collect blood an hour before an anticipated rise in temperature. To increase chances of isolating the pathogen at least 2 specimens should be collected at different times.

VOLUME

Adults 5-10ml blood and in children 2-3 ml should be drawn. Blood to media ratio is 1:10. Insert needle through the rubber liner &dispense.(in Brain heart infusion broth or whatever medium is available).

Blood culture bottles

C.S.F COLLECTION & TRANSPORT

From arachnoid space,using a wide-bore needle between L4 & L5.(Aseptic technique)

CSF is allowed to drip into two separate dry,sterile containers.( for C/S,other invest.) A Ventricular puncture is sometimes performed to collect CSF in infants. Notify lab before performing,L.P so that specimen is received&Exam immediately.

TRANSPORT OF CSF- AT 37oC

DELIVER IMMEDITALY BY KEEPING NEAR BODY OR IN TRANSPORT MEDIUM LIKE TRANSGROW OR TRANSISOLATE ( TIM) IMMEDIATE PROCESSING IN LAB OR KEEP IN INCUBATOR at 37oC

Anaerobic blood culture

Take same precautions as for aerobic culturing. Dispense 5 ml blood in thioglycollate medium(50 ml broth) or brain heart infusion broth.

Urine Collection

Morning urine specimen to be sent. MAINTAIN COLD CHAIN. OTHERWISE RANDOM SPECIMEN.
Sterile, dry, wide neck, leak-proof container.

Urine Collection

Instruct the patient to clean the area. 10-20 ml clean catch , MSU specimen. In Catheterised patient, aspirate with needle b/w tip&bag.

Urine sample storage

Immediate delivery not possible refrigerate at 4-6oC OR boric acid as preservative.which allows bacteria to remain viable without multiplying.

Faeces collection&transport

Patient to be given clean, dry, disinfectant free bedpan/wide-necked container (non sterile). Avoid contamination of faeces with urine. Separate worms, tape worms and segments from faeces if any. Gross examination information to be provided.

TRANSPORT MEDIA FOR FAECES

Cary-Blair media for Salmonella,Shigella,Vibrio and Yersenia(for 48 hrs). Cholera 1 ml specimen in 10 ml of alkaline peptone water(8 hrs). Worms, tapeworms segments in normal saline.

Cotainer for faeces &urine

Collection of pus,ulcer & skin specimens

Pus from abscess is best collected at the time when: Abscess is incised,drained or after it has ruptured naturally. Avoid contamination from skin commensals.
Specimen should be collected before Antiseptic dressing is applied.

Collection of Specimen from ulcers

Aspirate from a drainage tube up to 5ml of pus.

If pus is not being discharged : use a cottonwool swab to collect sample.Immerse swab in a container of Amies transport medium.

When the tissue is deeply ulcerated/necrotic:Aspiration is done from the side-wall of ulcer.

Collection of Serous fluid/exudates

Fluid from pustules,buboes & blisters is aspirated with a syringe. Serous fluid from skin ulcers ,papillomas may contain treponemes.

A drop of exudate is collected directly on cover glass & inverted on a clean slide. Immediately the specimen is dispatched for dark-field microscopy.

Transport of pus/ulcer specimens

Specimen is collected using a sterile cottonwool swab & inserted in a container of Amies transport medium. Aspirated specimen is transferred to a sterile, leak-proof container & sealed. A smear is made on a clean slide for Gramstaining.

Urogenital specimens

Collected by a medical officer or an experienced nurse. Amies medium, for transporting urethral,cervical,and vaginal swabs within 6 hrs. If cotton swab used then immediately deliver to the lab & proceed without delay.

Take urethral specimens at least 1 hour after the patient has urinated. Collect pus directly on a swab. If no discharge is evident, the male urethra is stripped towards the orifice to evacuate exudate. If no exudate is obtained, insert a thin swab [special urethral swabs if available] 2-3 cm into the urethra and gently scrape the mucosa by rotating the swab for 510 seconds. In women, massage the urethra against the pubic symphysis and use the same technique as for men.

Endocervical Swabs: With the patient in lithotomy position, expose the cervix using a vaginal speculum. Moisten the speculum with warm water since many lubricants contain antibacterial substances which may inhibit growth of gonoccoci. Collect the following specimens

According to following instructions Remove any mucous plug from the external os using a cotton ball. Insert a Transport medium swab into the endocervix. Rotate it gently and move it from side for 30 seconds before removal. Transport this swab to laboratory at room temperature 20-22O C ,taking care that it should get cultured on MTM plates within 12 hours preferably within 6 hours if amies transport media is used. Inoculate the swab directly on the labeled MTM plate only if the plate is provided at the collection site. Discard this swab. Using sterile swab, collect a second specimen from the endocervix as above and make a smear on the slide labelled CX let it air dry. Discard the swab.

Fluids and effusions

Synovial, pleural, pericardial, ascitic & hydrocele fluids. Collected by a medical officer aseptically. Transport immediately to the lab. collect into two portions ,one with anticoagulant 3%sodium citrate 1ml and 9ml fluid for cell count,protein estimation,microscopy,and culture Other 5 ml in screw capped container

Fluids and effusions

Alternate method: Dispense 5-10 ml fluid in blood culture bottle( aerobic or anaerobic), transport not over 37 C or in direct light. Transport to lab without delay.

12.FLUID / EFFUSIONS Physical Examination

Centrifuge 10 ml sediment for culture Gram and Z-N

OR
When positive

5-10 ml

Semen

Give pt. a clean,dry,leak proof container. Transport in a plastic bag and kept in the pocket to maintain the temperature.

Collection for ANAEROBES

APPROPRIATE SPECIMEN PUS IN SYRINGE WITHOUT AIRCOLUMN OR IN ROBERTSONS COOKED MEAT MEDIUM BROTH NOTE SWABS ARE UNFIT

Collection Procedures

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