PROCEDURE FOR REPORTINNG OF WORKLOAD, QUALITY CONTROL AND

INVENTORY CONTROL

WORKLOAD:

 It is the responsibility of the medical technologist to record all laboratory results every
day to include referrals or send out specimens.
 Results are sorted out into 3 sections namely Hematology, Chemistry and Clinical
Microscopy. A referral or send out record book is also provided.

 Mostly statistical and financial report should be made stating the total number of
specimens received, total specimens examined, total send outs and the reason for send
outs, re no reagents, equipment failure or not done.

 Annual reports should also be made stating same as above and be submitted to the
pathologist and management on the first week of January of the following year.

QUALITY CONTROL:

Laboratory staff must be able to recognize whenever a test fails to perform as expected.
This is accomplished by complying with the following requirements.

 The entire testing process must be continually monitored to ensure that laboratory errors
are promptly identified and corrected before a laboratory result is reported.

 Written procedures must be reviewed and signed by the pathologist on an annual basis
and placed in a lab manual available to staff at each testing site.

 The procedure must adhere to all requirements specified by the manufacturer of the test
kit or control.

 All test materials (controls, reagents, and supplies) must be stored in accordance with
the conditions specified in the procedure.

 Staff members may not exchange reagents from one kit with another. Likewise, do not
use expired materials for patient testing.

1
 Corrective actions must be initiated whenever the laboratory suspects that errors in laboratory
testing have occurred. This includes the pre-analytic, analytic, and post-analytic phases of
testing. Specific instances which require corrective include, but are limited to, the following

Errors in specimen collection or handling re indentified

Errors were detected in proficiency test results.
Poor methods of performance
Errors were detected in proficiency test results.
Errors in laboratory reporting are identified.

INVENTORY OF LAB SUPPLIES AND REAGENTS:

This procedure shall apply to all personnel who order any reagents for use in the laboratory.

Laboratory stuff shall ensure that there will be an establish inventory management program to ensure
that: *supplies and reagents are always available when need; *high quality reagents are obtained at an
appropriate cost; *reagents and supplies are not lost to improper storage or kept and used beyond
expiration and prevent overstocking which leads to wastage of supplies.

1. LGU shall ensure annual budget requirements for the purchase of needed supplies and reagents for
the laboratory that an uninterrupted supply of consumables and/or services is available to perform
all quality laboratory functions.
2. The laboratory has a documented procedure for ordering, receiving, documents, evaluating and
storting all consumables supplies.

EQUIPMENTS INVENTORY

All equipment information shall be entered into an equipment inventory record containing the
following information:

 Name of equipment location of the laboratory

Model and serial numbers manufacturer and vendor contract information
 Date of purchase warranty
 Date of commissioning

Inventory number shall be attracted to the equipment for ease of reference. Equipment not in
use shall be kept in a store or lockable cupboard and not in the main working areas.

2
PROCEDURE IN THE ACTUAL PERFORMANCE OF EQAP ACTIVITIES

Secure a NEQAS Registration Form

Accomplish the form Submit to NRL (directly or online)

Pay corresponding participation fee to NRL (directly or bank transfer

NRL acknowledge receipt of application

NRL sends samples to participants

Participants tests samples

Submit results to NRL

NRL provides results to participants

NRL issues certificate of participation to participants

3
 PROCEDURE ON REFERRAL AND OUTSOURCING OF EXAMINATIONS

To ensure specimen integrity on referral and transport adherence to the following steps should be
followed.

o All specimens sent to referral laboratories shall be collected and handled according to the
precise requirements of the policy of the referral laboratory. These include specimen
temperature, transport time and separation of cell from serum or plasma and slide
fixation.

o Detailed patient information must be provided in all cases.

oPatient confidentiality must be maintained at all times.

oSpecimens must be packaged and preserved appropriately.

oAppropriate temperature and transit items must be maintained.

o Packaging of specimens for shipment must be designed to minimize breakage

o Specimens shall be labeled with a biohazard label during transportation

oAll transactions on referrals will be documented and recorded in the appropriate logbook.

PRODECURES FOR REPORTING AND ANALYSIS OF INCIDENTS AND ADHERE

EVENTS

o All laboratory incidents and adverse events shall be documented and reported to the
management and to the pathologist. There shall be contact numbers provided for this
purpose and checked periodically to ensure that they are functional.

o It is the duty of the laboratory staff to manage any adhere events /accidents and to make a
verbal and written report and to notify promptly the head of laboratory and the
management.

o First Aid Kits should be available and maintained for treatment of minor injuries or for
short-term emergency treatment before getting medical assistance.

4
 IDENTIFICATION OF SPECIMEN:

 Every specimen received for testing shall be numbered, listed in an accession book or
otherwise marked so that it may be identified definitely and related to the patient, and the
submitting member of the healing arts or the referring laboratory

 An appropriate dated record of its receipt, disposition and examination, findings obtained,
and charges assigned shall be made and kept on file for the minimum period required by
statue. The records shall be available for inspection by authorized representatives of the
Department

 Contents of records. Each clinical laboratory shall have a readily available record
indicating the daily accession of specimens containing the following information:

(1) The laboratory number identifying the specimen.

(2) The identification of the person from whom the specimen was taken.

 Specimens received by mail shall be accompanied by the name and address of the patient
from whom the specimen was taken.
(3) The name and address of the licensed practitioner of the healing arts or other
Authorized person or clinical laboratory who submitted the specimen.
Hospitals may follow their normal procedures associated with requests and
records.
(4) The date and hour the specimen was collected by the lab or other authorized
person. In the laboratory.
(5) The date and hour the specimen was received in the laboratory and date test
was completed.
(6) The condition of the specimen when received that is, broken, leaked,
hemolyzed, turbid, satisfactory and so forth
(7) The analysis performed.
(8) The result of the laboratory test.
(9) The date a required report was released.

5
 VENIPUNCTURE PROCEDURE

1. Have the patient sit in a comfortable, study chair with his/her arm supported on a
table or chair arm for easy access. A patient should never stand or sit on a high stool
during the process of blood collection. The phlebotomist should always be prepared
for the occasional patient has an adverse reaction to phlebotomy.

2. Place the arm in a downward position to prevent back-flow. Apply a tourniquet three
to four inches above the puncture site, just tight enough to be slightly uncomfortable.
Ask the patient to make a fist. Do not allow the patient to make a very tight first since
this can falsely elevate the potassium level. This makes the veins more palpable.

Do not leave the tourniquet on for more than 1 minute. It may be necessary to release
the tourniquet after vein selection and to reapply it immediately prior to the puncture.

3. Check both arms. The three main veins are the cephalic, median cubital, and basilic.
Generally, the median cubital is the one of choice because it is well-anchored in
tissue and will not roll or move when the needle punctures it. The median basilic, at
the inner edge of the arm, may have tendency to roll and is near a main artery and
nerve. This part of the arm is very ender. Thecephalic vein also has a tendency to roll
and the skin over it is often tough.

4. Using the index finger, palpate the arm, feeling for the best vein. It should feel similar
to an elastic tube.

5. If a vein cannot be found try the following suggestions.

a. Gently pat the site to enlarge the veins
b. Massage the arm
c. Wrap the arm in a warm towel
d. Check both arms. Always select the most suitable vein for puncture.

6
 VENIPUNCTURE PROCEDURE

FILLING OF TUBES:

a. If a syringe is used, blood will begin to flow as soon as the needle enters the vein. If using a
syringe, do not pull back too hard on the plunger; this may hemolyze the cells, collapse the
vein, or pull the wall of the vein over the bevel, stopping the blood flow. Keep the needle
steady; do not push or pull out. Continue pulling plunger back gently until enough blood is
collected to fill all needed tubes.

b. If a vacutainer is being used, as soon as the needle in the vein, push the tube firmly
butcarefully into the holder centered onto the back of the needle until resistance is felt. If
the vein has been located, blood flow will begin at the resistance point. Keep the needle
steady. If collecting multiple samples, wait until the vacuum is exhausted and blood flow
ceases. Gently remove the tube from the holder, keeping the needle steady, and place the
tube into the holder.

The proper order of draw is:

i. Blood culture bottles

ii. Non-additive red or blue if any coagulation test other than PT/PTT ordered (invert
3-4 times)
iii. Blue top- must be at least 90%
iv. Gold top (invert 5 times)
v. Green top (invert 8-10 times)
vi. EDTA pink and/or purple (invert 8-10 times)
vii. Any other tubes

7
 Blood Collection by Finger Prick

Materials and Equipment:

Disposable gloves cotton wool

Sharps disposal containers 100% household bleach
Alcohol swabs
Sterile bloods lancets
Cotton wool
10% household bleach
Capillary tubes or disposable Pasteur pipettes

Procedure:

1. Explain to the client about the procedure.

2. Wear disposable gloves and use aseptic for blood collection

3. Position the client; he/she should sit in a chair and hyperextend his/her arm.
4. The best locations for a finger prick are the third and fourth fingers of the hand.
Do not use the tip or the centre of the finger. Avoid the side of the finger.
5. Avoid puncturing a finger that is cold or blue, swollen, scarred or covered with a rash.
6. Warm the finger by wrapping a warm cloth during cold or by rubbing the finger with
your hand.
7. Use a sterile lancet. Full skin penetration by the tip of the lancet should be
accomplished in order obtain adequate blood flow for collection.
8. Make the skin puncture just off the centre of the finger pad.
9. With dry and clean cotton wool, wipe off the first drop of blood.
10. Gently massage the finger to allow a drop to form at the punctured site. Collect
sufficient quantities of blood for the technique in question using the recommended
equipment.
11. Have the patient hold a small ball of dry cotton wool over the puncture site for a few
minutes to stop the bleeding.

8
 VENIPUNCTURE PROCEDURE

TROUBLE SHOOTING GUIDELINES:

IF AN INCOMPLETE COLLECTION OR NO BLOOD IS OBTAINED:

Change the position of the needle. Move it forward (it may not be in the lumen)

 Or move it backward (it may have penetrated too far).

 Adjust the angle (the bevel may against the vein wall
 Loosen the tourniquet. It may be obstructing blood flow.
 Try another tube. Use a smaller tube with less vacuum. There may be no vacuum in the
tube being used,
 Re-anchor the vein. Veins sometimes roll away from the point of the needle and puncture
site.
 Have the patient make a fist and flex the arm, which helps engorge muscles to fill veins.
 Pre-warm the region of the vein to reduce vasoconstriction and increase blood flow.
 Have the patient drink fluids if dehydrated.

9
 MANUAL WHITE BLOOD CELL COUNT:

PRINCIPLE:

Whole blood is diluted with a 3% acetic acid solution, which hemolyzes mature erythrocytes and
facilitates leukocyte counting. The standard dilution for leukocyte counts is 1; 20. This dilution is
prepared using the leukocyte Unopette system. The dilution is mixed well and incubated to
permit lysis of the erythrocytes. Following the incubation period, the dilution is mounted on a
hamacytometer. The cells are allowed to settle and then are counted in specific areas of the
hemacytometer chamber under the microscope. The number of leukocytes is calculated per m L
(x10 to the 9th power/ L of blood.

PROCEDURE:

(1) Draw well-mixed capillary or venous blood exactly to the 0.5 mark in a blood cell
diluting pipet. This blood column must be free of air bubbles.

(2) Wipe the excess blood from the outside of the pipet to avoid transfer of cells to the
diluting fluid. Take care not to touch the tip of the pipet with the gauze.

(3) Immediately draw diluting fluid to the “11” mark while routing the pipet between the
thumb and forefinger to mix the specimen and diluent. Hold the pipet upright to prevent
air bonules in the bulb.

(4) Mix the contents of the pipet for 3-5 minutes to ensure even distribution of cells. Expel
unmixed and relatively cell-free fluid from the capillary portion of the pipet (usually 4
drops).

(5) Place the forefinger over the top (short end) of the pipet, hold the pipet at a 450 angle,
and touch the pipet tip to the junction of the cover glass and the counting chamber.

10
 MANUAL WHITE BLOOD CELL COUNT

(6) Allow the mixture to flow under the cover glass until the chamber is completely charged.
Similarly, fill the opposite chamber of the hemacytometer. If the mixture overflows into
the moat or air bubbles occur, clean and dry the chambers remix the contents of the pipet,
and refill both chambers.

(7) Allow the cells to settle for about 3 minutes. Under low-power magnification and
reduced light, focus on the ruled area and observe for even distribution of cells.

(8) Count the white cells in the four 1 sq mm corner areas corresponding to those marked A,
B, C, and D of Figure 5-1 in each of two chamber.

(9) Count all the white cells lying within the square and those touching the upper and right
hand center lines. The white cells that touch the left-hand and bottom lines are not to be
counted. In each of the four areas, conduct the count as indicated by the “snake-like” line
in.

Figure 5-1. A variation of more than 10 cells between any of the four areas counted or a
variation of more than 20 cells between sides of the hemocytometer indicate uneven
distribution and require that the procedure repeated.

11
 (1) Routinely blood is drawn to the 0.5 mark and diluted to the 11 mark with WBC fluid. All
the blood is washed into the bulb of the pipet (which has a volume of 10). Therefore, 0.5
volumes of blood are contained in 10 volumes of diluting fluid. The resulting dilution is
1:20. (These figures are arbitrary and refer strictly to dilution and not specific volumetric
measurements.)

(2) The depth of the counting chamber is 0.1 mm and the area counted is 4 sq mm (4 squares
are counted, each with an area of 10 sq. mm therefore, 4 x 10 sq mm = a total of 4 sq
mm.) The volume counted is: area x depth = volume. Four sq mm x 0.1 mm = 0.4 cu mm.

(3) The formula is as follows:

WBCs per cu mm = Average number of chamber (2) WBCs counted x dilution (20)
Volume (0.4)

MANUAL WHITE BLOOD CELL COUNT:

Sources of Error:

(1) Improper collection of blood specimens causes variable results.

(2) Wet or dirty pipets
(3) Poor condition or inaccurate calibration of pipets. Pipets must be in good condition and
calibrated to have maximum error of + 1 percent.
(4) Poor pipeting technique pauses high or low counts. Poor pipeting technique includes:

(a) Undershooting desired line with blood or diluting fluid.

(b) Overshooting desired line with blood or diluting fluid.
(c) Air bubbles in the column on bulb.
(d) Failure to wipe tip free of blood.
(e) Too slow manipulation following the withdrawal of the specimen thus, allowing some
of the blood specimen to coagulate.
(f) Failure to mix the blood and diluents properly.

12
 MANUAL WHITE BLOOD CELL COUNT:

(5) Failure to expel 2 or 3 drops in the pipet tips before charging the hemacytometer.
(6) Over filling the chamber of the hemacytometer, which causes erroneously high counts?
(7) Wet or dirty cover glasses and hemacytometer.
(8) Uneven distribution of cells in the counting chamber causes erroneous results.
(9) Inaccuracy or careless in marking counts to +15 percent.
(10) Diluent that which is cloudy or contains debris.
(11) Failure to mix anticoagualated blood thoroughly before use.

Normal Values WBC count:

(1) Adults (both sexes): 4,500- 11,500 WBCs per cu mm.

(2) Childhood: 6, 00-14,000 WBCs cu mm.
(3) Birth: 9,000-30,000 per cu mm.

PERIPHERAL BLOOD SMEAR PREPARATION

Procedure:
 Place a 1” x 3” glass microscope slide with a frosted end on a flat surface (usually the
counter top of a laboratory bench).
 Attach a label on the slide or write the patient name, identification number, and
preparation on the frosted surface.
 Place a 2-3 mm drop of blood approximately 1/4 “frosted slide, using a wooden
applicator stick or glass capillary tube.
 Hold the slide by the narrow slide between the thumb and forefinger of one hand at the
end farthest from the frosted end.

Peripheral Blood Smear Preparation

Procedure:

 Grasp second slide (“spreader slide”) between the thumb and forefinger of the other hand
at the frosted end.
 Place the edge of the spreader slide on the lower slide in front of blood (slide farthest
from the frosted end.

13
  Pull the spreader slide toward the frosted end until it touches the drop of blood. Permit
the blood to spread by capillary motion until it almost reaches the edges of the spreader
slide.
 Push the spreader slide forward at a 30° angle with a rapid, even motion. Let the weight
of the slide do the work.

Placing a small drop or venous blood on a glass microscope

Step 1 slide, using a glass capillary pipette. A wooden applicator stick
can also be used for the purpose.
A spreader slide has been positioned at an angle and slowly
Step 2 drawn toward the drop or blood.
The spreader slide has been brought in contact with the drop of
Step 3 blood and is being drawn away. Note layer of blood at the edge
of the spreader slide.
The speaker slide is further pulled out, leaving a thin layer of
Step 4 blood behind.
The blood smear is nearly complete.
Step 5
End result. A glass slide with a well-formed blood film. After
Step 6 drying for about 10 minutes, the slide can be stained manually
or placed on an automated slide stainer.

Fig. 1. Wedge slide technique for preparation of a peripheral blood smear.

A well-made peripheral smear is thick at the frosted end and becomes progressively thinner
toward the opposite end. The “zone of morphology” (area of optimal thickness for light
microscopic examination) should be at least 2 cm in length. The smear should occupy the
central area of the slide and be margin-free at the edges. (Fig. 2).

STAINNING PROCEDURE:

1. Fix the film by dipping for 2 seconds in absolute methanol. Allow dying
2. Stain using Leishman by flooding the slide for 1 to 3 minutes.
3. Dilute the stained slide with buff (ph 6.8) for 4 to 7 minutes.
4. Using a pair of forceps, hold one end of the slide and rinse with water to remove excess
stain.
5. Keep the slides on the rack in a standing position and allow drying for 5 minutes before
doing different counting.

14
D. Analysis

Procedure

1. Do the differential blood count.

2. Report the results on the Different Blood count and HCT worksheet.

Characteristics of a Good Smear

 Good smear is tongue shaped with a smooth tall.

 Does not cover the entire area of the slide.
 Have both thick and thin areas with gradual transition.
 Does not contain any lines or holes.

MANUAL WHITE CELL DIFFERENTIAL COUNT and PLATELET:

PRINCIPLE:
The differential white cell count is performed to determine the relative number of each
type of white cell present in the blood. This provides valuable information concerning
infections and other disease processes. At the same time, a study of red cell, white cell,
and platelet morphology is performed. Performing the differential smear to be used as a
double-check of the white cell count and platelet count.

15
II. Specimen: Whole blood from a capillary or EDTA at least ¾ full.

III. REAGENTS AND EQUIPMENT

a. Microscope
b. Differentiate Cell Counter
c. Stain: Leishman or Giemsa Stain

IV. QUALITY CONTROL

1. The RBCs should appear buff pink to orange

2. WBCs should have a blue nucleaus with a lighter- staining cytoplasm.

DIFFERENTIAL COUNT PROCEDURE:

1. Stain the with appropriate stain and place the slide on the microscope stage with smear
side up and focus using the low power objective and low light.
2. Scan the blood smear, nothing any unusual or irregular cells or rouleaux formation
3.
4.
5. Carefully change to the oil immersion (100x), focus and increase light intensity as
needed.
6. Begin in the thin area of smear
7. Scan the slide in figure” 5
8. Counts. :

A. White blood cell counts

(1) For WBC counts less than 20x109 WBC/L, count and classify 100 cells.
(2) For WBC COUNTS of 20% to 50% x 109 WBC/L, count and classify 300 cells.
(3) If there are an abnormal percentage of cells in the differential count, classify 200
cells.
(a) More lymphocytes than neutrophils (except in children).
(b) Over 11% monocytes
(c) Over 10% eosinophils
(d) Over 2% basophils

B. Nucleated red blood cells (NRBCs):

(1) Count number of NRBCS PER 100 WBCs on a separate counter; report.
(2) Recalculate the WBC count if the NRBC count is greater than 5NRBCs per
100 WBCs.

9. Report any abnormalities of white cells

16
 C. PLATAELET ESTIMATE
(1) Scan the thin area, using the oil immersion lens.
(2) Observe 10 fields, counting the platelets in each field, observing granulation
(3) Determine the average number of platelets observed per oil immersion field
( OIF)
(4) Report the platelet estimate and any abnormal morphology.

Normal Count: 150, 00-350, 00/mm3

D. WBC ESTIMATE:
1. Scan the thin area, using the 50x oil immersion lens.
2. Observe 10 fields, counting all WBCs in each field
3. Average the number of WBCs seen per oil immersion field (OIF)
4. Document performance of WBC estimate.
5. Report any discrepancies to pathologist.

E. RBC MORPHOLOGY
1. Scan the thin area, using the oil immersion lens.
2. Observe 10 fields.
3. Report RBC size and shape. Use the nucleaus of a typical small lymphocyte as
a comparison for normal size.
4. Report any alterations in colour, the amount of hemoglobin, or inclusions
 Save all differential slides for 7 days.

Normal Values for Total WBC and Different Count:

Total WBC: 4,500= 10,000

 Bands or stabs: 3-5%

 Granulocytes (or polymorphonuclears)
o Neatophills (or segs): 50-70% relative value ( 2500-7000 absolute
value)

17
 o Eosinophils: 1-3 relative value (100-300 absolute value)\
o Basophils: 0.4% relative value ( 40-100 absolute value)
Agranoculytes (or mononuclears)
o Lymphocytes: 25-35% relative value (1700-3500 absolute value)
o Moncytes: 4-6% relative value (200-600 absolute value

Manual Platelet Count

(Direct Method)

Principle:

Whole blood is dilluted with a 1% ammonium oxalate solution. The isotonic balance
of the diluents is such that all erythrocytes are lysed while the leukocytes, platelets
anad reticulocytes remain intact. The standard dilution for platelet counts is 1;100.
This dilution is prepared using the leukocyte/platelet. Unopette system. The dilution
is mixed well and incubated to permit lysis of the erythrocytes. Following the
incubation period, the dilution is mounted on a hemacytometer chamber under the
microscope. The number of platelets is calculated per m L(x109 /L) of blood.

Reagents and Equipment

 Two leukocyte/platelet Unopette reservoirs; each containing 1.98mL of the
following diluents:

Ammonium oxalate 11.45 g

Sorensen’s phosphate buffer 1.0 g
Thimerosal 0.1 g
QS with distilled water to 1 liter
 Two Unopette capillary pipets, 20yl 4. Petri dish filter
 Hemacytometer with cover glass 5. Microscope
 Petri dish with filter paper 6. Hand Counter

Commercial quality control materials with established control limits should be run periodically.
The frequency is detrmined by each laboratory’s workload. For instance, quality control material
may be run at the beginning of each eight-hour shift.

SPECIMEN:

Whole blood, anticoagulated with EDTA, or free-following capillary blood may be used.

18
 Manual Platelet Count
(Direct method)

PRINCIPLE
Procedure:

1. Prepare two leukocyte/Platelet Unoppettes as follows:

Using the proactive shield on the capillary pipet, puncture the diaphragm as follows:
1) Place reservoir on a flat surface. Grasping the reservoir in one hand, take the pipet
assembly in the other hand and push the tip of the pipet shield firmly through the
diaphragm in the neck of the reservoir then remove.
2) Remove the shield from the pipet assembly with a twist and fill the capillary pipet with
whole blood. Transfer the whole blood to reservoir as follows:
1) Wipe excess blood from the outside of the capillary pipet, making certain that no
blood is removed from the capillary bore.
2) Squeeze the reservoir slightly to force out some air. Maintain pressure on the
reservoir.
3) Cover opening of overflow chamber of the pipet your index finger and seat the pipet
securely in the reservoir neck.
4) Release pressure on the reservoir. Then remove your finger from the piper opening.
Negative pressure with draw the blood into the diluents.
5) Squeeze the reservoir gently two or three times to rinse the capillary bore, forcing
diluents into, but not out of, the overflow chamber, releasing pressure each time to
return the mixture to the reservoir.
6) Place your index finger over the pipet opening and gently invert several times to
thoroughly mix the blood with diluent.
7) Let stand for 10 minutes to allow erythrocytes to hemolyze.

Normal Count : 150,000 – 350,000/mm3

INDIRECT PLATELET COUNT:

 Make a good blood smear from a free flowing skin puncture or from a various blood
taken from EDTA.
 Dry into the air and stain with a wright stain.
 Using an oil immersion objective, count the number of platelets seen per 100x oil
immersion field.

19
  10 oil immersion fields are counted and the results averaged. (This accounts for even
dispersal of platelets in the smear.
 Then the following formula is applied.

Estimated platelet count/yL = average count in 10 fields x 15,000.

HEMOGLOBIN TEST:

PRINCIPLE

Ferrycyanide oxidizes oxyhemoglobin to methmoglobin, and cyanide coverts methemoglobin to

cyanmethomoglobin (3). Absorbance measurements are made at 540 nm. The
CYANMETHOMOGLOBIN REAGENT contains a surface to promote rapid hemolysis and to
accelerate formulation of cyanmethomoglobin. The reaction is completed in 3 minutes.

PROCEDURE:

1. Place 2 Ml of CYAMETHOMOGLOBIN STANDARD in tube labeled standard.

2. Disperse 5.0 mL CYAMETHOMOGLOBIN REAGENT into tubes labeled Reagent
Blank, Control, Sample 1, etc.
3. Place 0.02 mL specimen into the appropriately labeled tube. Use delonized water as
specimen for Reagent Blank, Mix wel.
4. Allow the test samples to stand at room temperature (15-30°C) for at least 3 minutes.
5. Adjust instrument to zero absorbance at 540 nm using Reagent Blank.
6. Read and record absorbance values for Standard, Controls and Unknowns.

NOTE: For a direct read-out instrument, set read-out concentration value of the Standard and
read the unknown concentrations directly.

STABILITY OF FINAL REACTION

Cyanmethemoglobin appears quite stable, However, the test samples be read within an hour
before evaporation of the reaction solutions becomes significant.

NORMAL VAUES: Conventional Units (g/dL) SI Units (g/L)

20
Adult Males: 13.5-17.5 135 – 175
Adult Females: 12.0 – 16.0 120 – 160

HEMATOCRIT

1. Using a fresh lancet lance the tip of your middle finger (on the side) to stimulate
bleeding. Raising the opposite arm will promote increased blood flow to the bleeding
finger. Be sure to have your opposite hand gloved!

PROCEDURE

STANDARD PRECUTION:

Patient’s specimen and all materials coming into contact with them should be handled as if
capable of transmitting infections and disposed of with precautions. Gloves should be worn
when handling all specimens.

1. Place both capillary tubes in the centrifuge across from each other. The sealed end should
be on the outside, as the centrifugal forces press the cells toward the outside of the
centrifuge.
2. Place the lid on the centrifuge head. Most centrifuges have a special spin-on head to
prevent the capillary tubes from flying out of the head during the centrifugation.
3. Centrifuge the capillary tubes for 5 minutes at a minimum of 15, 000 g.
4. Remove the capillary tubes after spinning has stopped. To read the micro hematocrit,
place the tube at the right edge of the reader card (see Figure 1) with the top of the
plasma line at the 300% mark. Slide the tube to the left until the top of the day sealant is
at the bottom line (0). Check that the top of the plasma is at the 100% line, the bottom of
the red cell column is at the 0 line, and the capillary tube is parallel to the edge of the
reading card.
5. Carefully find the point where the top of the red column crosses the line of the reader
card.

Dot includes the gray buffy coat just above the red cell column as part of the reading. The
identified line corresponds to the micro hematocrit value. It may be to use a magnifying glass
to accurately identify the top of the red column.

21
 6. Repeat the reading process for the second capillary tube. The results should agree within
two percentage points. The reported value is the average of those values. If the two
values differ by greater than 3 points, the entire procedure should be repeated.

Normal Hematocrit for Adult:

Females: 37-47% (Ave. 45%)
Males: 42-52% (Ave. 47%)

NOTE: Microhematorit Reader. Abnormal values may suggest either deficiencies

(anemia) or excesses (erythrocytis or polycythemia)

BUILDING TIME: (Dukes Method)

Principle:
Determination of bleeding time measures the ability of small blood vessels to control
bleeding after injury. This depends on platelet plug formation, vasoconstriction and
coagulation. This test aids in the diagnosis of a certain bleeding diseases and provides a
safeguard in cased of proposed surgery.

Materials:

Disposable lancet/pricker
Filter paper, cut into pieces.
Watch or stop watch.

Procedure:

 Clean the earlobe or the tip of the finger with 75% alcohol and allow dying.
 Make a relatively deep puncture with a sterile blood lancet and the stopwatch.
 There should be a free flowing blood. Avoid milking the finger or applying pressure
 Using the circulator filter, blot the blood every 30 seconds. Do not allow the filter
paper to touch the wound.
 When bleeding ceases stop the stopwatch and record the bleeding time.

CLOTING TIME: (Slide Method)

PRINCIPLE:

22
A blood sample is obtained to determine the clotting process assessed in vitro by measuring the
time from taking the blood sample to the appearance of the first clot.

Materials: Glass slide

Disposable Lanet/Pricker
Stopwatch/Watch

Procedure:

 Puncture the fingertips with disposable lancet after cleaning

 Collect 2 separate drops of blood on the glass slide.
 Start the stopwatch, and check every 30 seconds by disturbing the blood on the glass slide
with the tip of the lancet.
 When you observe fibrin shred beginning to develop. Then coagulation is developing, a
record type.
 Then coagulation is developing re, record time.

RETICULOCYTE COUNT

PRINCIPLE:

Nonnucleated immature erythrocytes contain nuclear remnants of RNA and the cell is
known as a reticulocyte. To detect the presence of this RNA, the red cells must be stained
while they are living. This process is called supravital staining. With supravital staining,
the RNA appears as a reticulum within the red cell.

Reagent.

(1) New Methylene Blue Solution. Dissolve 0.5 grams of new Methylene Blue, 1.4 grams
of potassium oxalate, and 0.8 grams of sodium chloride in distilled water. Dilute to
100 ml. Filter before use.
(2) Brilliant Cresyl Blue Solution. Dissolve 1.0 grams of brilliant cresyl blue 99 ml of .85
percent sodium chloride, Filter before use.

Procedure:
(1) Place 3 or 4 drops of new methylene blue and 3 or 4 drops of blood (venous or capillary)
in a small test tube.
(2) Mix the tube contents and allow to stand for a minimum if 15 minutes. This allows the
reticulocytes adequate time to take up the stain.

23
 (3) At the end of 15 minutes, mix the contents of the tube well.
(4) Place a small drop of the mixture on a clean glass slide and prepare a thin smear.
(5) Counterstain with Wright’s stain, if desired
(6) Allow smear to air-dry.
(7) Place the slide on the microscope stage and, using the low power objective, locate the
thin portion of the smear in which the red cells are evenly distributed and are not
touching each other.
(8) Switch to oil immersion magnification and count the number of reticulocytes in 5 fields
of 200 RBCs.

d. Calculation

Number of reticulocytes counted

% reticulocytes =
10

RETICULYTE COUNT

Sources of Error:
(1) Equal volumes of blood and stain give optimum staining conditions. An excess of blood
causes the reticulum to understain. An excess of stain usually obscure the reticulum.
(2) Crenated erythrocytes and rouleaux formation male an accurate count difficult to perform.
24
(3) Stain precipitated on erythrocytes causes them to appear as reticulocytes
(4) Dirty slides cause uneven spreading.
(5) The dye solution should have adequate time to penetrate the cell and stain the reticulum.

Discussion:

(1) Reticulocytes are nonnucleated erythrocytes that exhibit blue reticulum stands within
their cytoplasm when stained supravitally. When stained only with Wright’s stain, they
are buff-pink in color and larger and darker than erythrocytes.

(2) Reticulocytes serve as an index of the activity of the bone marrow in blood regeneration.
As such, these counts are of value in following anti-anemia therapy. Satisfactory response
to therapy is evidenced by an increase of reticulocytes in the peripheral blood. Increased
reticulocyte counts also occur whenever there is rapid bone marrow activity as in
leukemia or blood regeneration associated with hemorrhage or hemolysis. Decreased
reticulocyte counts occur in conditions in which the bone marrow is not producing
adequate red blood cells, such as aplastic anemia.

(3) Several methods for staining and counting reticulocytes are in common use. Compared to
the use of alcohol solutions of dye, methods employing saline solutions of new methylene
blue can give slightly higher values for reticulocytes. For comparative studies, the same
method should be used throughout the work.

(4) Precipitated stain is often confused with reticulum but can be recognized by its presence
throughout the smear and apart from the red cells. Precipitation can be eliminated as a
source of error by frequently filtering the stain.

(5) An alternate method of counting reticulocytes utilizes the Miller disk that is placed inside
the microscope eyepiece. This disc consists of 2 squares as shown below in figure 5-2.
The area of the smaller square B, is a tenth that of square A. Therefore, if there are 40 red
cells in square A, there should be four red cells present in square B. When employing this
method to count reticulocytes, the red cells in square B are counted in successive fields
on the slide, until a total of 500 red cells have been counted. At the same time, the
reticulocytes in square A are enumerated. At the completion of the count, theoretically,
the reticulocytes obtained in this way are divided by 50, in order to obtain the present
reticulocytes present in the blood.

A 25
 B

Figure 5-2. Miller Disk

Normal Values:

(1) The birth. Two and one half to 60 percent, but falls to adult range by the end of second
week of life.
(2) Adults (both sexes). Five-sevenths to 1.5 percent.

ERYTHROCYTE SEDIMENTATION RATE

The ESR (erythrocyte sedimentation rate) measures the distance red blood cells will fall along
the length of a vertical tube over a given time period. A raised ESR reflects an increased
production of acute-phase proteins. Although it is a non-specific phenomenon, it is clinically
useful in certain chronic disorders, e.g. rheumatoid arthritis or tuberculosis, as an index of
progress of the disease.

26
A normal ESR does not include organic disease but, on the other hand, the vast majority of acute
or chronic infections and most neoplastic and degeneration diseases are associated with changes
in the plasma proteins which lead to an acceleration in sedimentation.

PRINCIPLE:

The recommended Westerngreen sedimentation tube is made from either glass or plastic, has a
length of about 30 cm and a bore of 2.5 mm. many of the tubes which are now available are
intended to be disposed of after use. Glass tubes can be re-used provided that they are adequately
cleaned by washing through with water, followed by alcohol, and then allowed to dry overnight.

Specially made racks with a scale graduated in mm from 0 to 140 are available and these have
adjustable leveling screws for holding the tubes in an exactly vertical position.

It is conventional to set up sedimentation rates at room temperature (18-25℃).

Sedimentation is normally accelerated as the temperature rises and, if the test is to carried out at
a higher ambient temperature, a normal range should be established for that temperature. The test
should be protected from direct sunlight and draughts, and it should never be set up near a
radiator or on a bench where there is vibration, e.g. from a centrifuge.

The test is performed on venous blood diluted accurately with 31.3 g/l trisodium citrate in the
proportion of one volume of citrate to four parts of blood. It should be carried out within two
hours of collecting the blood, though a delay of up to six hours is permission provided the blood
is kept at 4℃.

Blood in EDTA can also be used for up to 24 hours after collection provided that it is kept at 4℃
and diluted with trisodium citrate immediately before setting up the test.

ERYTHROCYTE SEDIMENTATION RATE

The ESR (erythrocyte sedimentation rate) measures the distance red blood cells will fall along
the length of a vertical tube over a given time period. A raised ESR reflects an increased
production of acute-phase proteins. Although it is a non-specific phenomenon, it is clinically
useful in certain chronic disorders, e.g. rheumatoid arthritis or tuberculosis, as an index of
progress of the disease.

27
A normal ESR does not exclude organic disease but, on the other hand, the vast majority of acute
or chronic infections and most neoplastic and degenerative diseases are associated with changes
in the plasma proteins which lead to an acceleration in sedimentation.

PRINCIPLE:

The recommended Westerngreen sedimentation tube is made from either glass or plastic, has a
length of about 30 cm and a bore of 2.5 mm. many of the tubes which are now available are
intended to be disposed of after use. Glass tubes can be re-used provided that they are adequately
cleaned by washing through with water, followed by alcohol, and then allowed to dry overnight.

Specially made racks with a scale graduated in mm from 0 to 140 are available and these have
adjustable leveling screws for holding the tubes in an exactly vertical position.

It is conventional to set up sedimentation rates at room temperature (18-25℃).

Sedimentation is normally accelerates as the temperature rises and, if the test is to carried out at a
higher ambient temperature, a normal range should be established for that temperature. The test
should be protected from direct sunlight and draughts, and it should never be set up near a
radiator or on a bench where there is vibration, e.g. from a centrifuge.

The test is performed on venous blood diluted accurately with 31.3 gl trisodium citrate in the
proportion of one volume of citrate to four parts of blood. It should be carried out within two
hours of collecting the blood, through a delay of up to six hours is permissible provided the
blood is kept at 4℃.

Blood in EDTA can also be used for up to 24 hours after collection provided that it is kept at 4℃
and diluted with trisodium citrate immediately before setting up the test.

ERYTHROCYTE SEDIMENTATION RATE

(Wintrobe Method)

PROCEDURE:

 Mix the blood well and draw the same sample into clean dry Westerngreen tube.

28
  Place the tube into the stand, taking care that the base is firmly positioned on the base pad
to prevent leakage. Adjust the rack so that the tube rests in an exactly vertical position.
 Leave undisturbed for 60 min.
 At the end of the hour read the height of clear plasma above the upper margin of the
column of sedimenting cells to the nearest millimeter.
 A poor delineation of the upper layer of red cells, so-called ‘stratisfied’ sedimentation,
has been attributed to the presence of my reticulocytes.
 Report this measurement as the ESR (Westerngreen) in units of mm in 1 hour.

Sources of Error

 Specimen older than specified time after collection and before testing (see above)
 Incorrect proportion of anticoaligulant
 Incorrect type of anticoagulant
 Hae molysed sample
 Contaminated sedimentation tubes
 Tubes tilted during sedimentation
 Test set up near central heating or direct sunshine
 Test set up adjacent to centrifuge or other instrument causing vibration

Failure to read at exactly one hour.

NORMAL VALUES:

Maless: 0-9 mm/h

Females: 0-15 mm/h

URINALYSIS

Procedure of urine sample collection:

Patient is given a clean bottle of 5 to 10 ml enclosed with a cap and informed to clean the
surrounding area with swab, and while voiding urine the person collects a small quantity of fresh

29
morning sample in that bottle, usually a midstream sample is collected and handed over the
enclosed bottle along with patient details on it to the laboratory or testing site within an hour of
collection.

Methods of urine analysis:

 Naked eye testing for Colour and cloudiness of the urine is noted through naked eye.
Yellow, amber, pale or red are the few colour types which can be noted and clear or
cloudy consistency and also the smell of urine is sensed which is of useful importance.
 Protein testing: take around 1ml of urine sample in 2 separate test tubes. Put one tube into
the how water bath, and leave the other at room temperature. After a few minutes, take
the test tube out of the water bath, and compare the heated and unheated urine. If the
heated sample is clouder, it contains protein.
 pH testing: Dip a piece of universal indicator paper into the urine. Quickly take it out, and
leave it for 30 seconds. Compare the new colour with the pH colour chart, and note the
pH
 Testing for glucose: this test is used to know whether there is any presence of glucose in
the urine, if yes then it can indicate disease conditions like Diabetes.
 Urine dipstick Test: it is a long strip which has a several squares of different colors on it.
Each square is used to interpret urinalysis. The strip is dipped completely in the urine
sample and color changes are noted after 5 minutes. The squares on the dipstick represent
the following components in the urine.
 Specific gravity, pH, protein in the urine glucose, ketone bodies, blood, leukocyte, nitrite,
bilirubin and urobilinogen.
 Microscopic evaluation: Cells, Cellular debris, Casts Bacteria and crystals or small
structures.

Microscopic Examination of Urine Sediment

As part of urinalysis, the urine sediment is centrifuged and examined microscopically

for crystals, casts, red blood cells, white blood cells, and bacteria or yeast. Because
examination of urinary sediment provides a direct sampling of urinary tract morphology, it

30
 provides important information useful for both diagnosis and prognosis. Microscopic
examination of urine sediment is usually performed in addition to routine procedures. This
examination requires a degree of skill acquired through practice under the immediate
supervision of an experienced technician. The specimen used for microscopic examination
should be as fresh as possible. Red cells and many formed solids tend to disintegrate upon
standing, particularly if the specimen is warm or alkaline.

How to Scan the Slide

Review the operating instructions for your microscope if you are unfamiliar with its
operation. Place the slide under the scope and begin the examination under lower power. Be
sure to use a low light source (adjust the iris and condenser). Too much light makes the
cellular and crystalline elements harder to see. Scan the slide under low power to locate areas
of interest. Look for casts just inside the perimeter of the cover slip. Count the numbers of
red cells and white cells in each and report the range of findings. If the field is covered with
cells, report as “TNTC” (Too numerous to count) or “packed”. Add to the report an estimate
of bacteria density, any casts seen and other structures noted, sample microscopic urinalysis
report 5-10 RBC/HPF, 15-25 WBC/HPF, few bacteria, occasional hyaline cast, no epithelial
cells.

A. Microscopic examination of urine sediment detects the presence and amounts of:
 Red blood cells
 White blood cells
 Bacteria and yeast
 Cast
 Epithelial cells
 Crystals

How to Scan the Slide

Urine sediment is assessed under a high power field (HPF) for the presence of red white blood
cells. Normally, there should be only an occasional red blood cell in the urine (2-3 per high

31
power field). Hematuria, the presence of abnormal numbers of red blood cells in the urine may
be due to:

 Glomerular disease Upper and lower urinary tract infections

 Tumors that erode any part of urinary tract Nephrotoxins
 Kidney trauma Traumatic catheterization
 Renal infarcts Passage of renal stones
 Acute tubular necrosis Physical stress

It is important that urine examined for bacteria, casts, crystals and epithelial cells. Urine stored in
the bladder is normally free of bacteria or yeast. However, the bacteria are commonly found in
urine specimens because of the abundant normal microbial flora of the vagina or external urinary
meatus and due to the ability of bacteria to multiple rapidly in urine standing at room
temperature. Bacteria noted on a microscopic examination should be interpreted in view of
clinical signs and symptoms of urinary tract infection. Diagnosis of bacteriuria in a patient with a
suspected urinary tract infection requires a urine cultured and sensitivity. A colony count may
also be done to determine if significant numbers of bacteria are present. Generally, more than
100,000 bacteria of one organism per millimeter of urine indicate a urinary tract infection. A
finding of multiple organisms usually reflects specimen contamination.

Casts are collections of protein, cells and debris that are formed in the tubules of the kidneys.
Cast width is described as narrow (one or two red blood cells in width), medium broad (three to
four red cells in width), and broad (five red blood cells in width) Casts that form in the collecting
tubules tend to be very broad. Broad casts usually indicate a significant reduction in the
functional capacity of the nephron and indicate severe renal damage or “end stage” renal disease.
A few hyaline casts are normal, but all other casts need to be evaluated. When cellular casts
remain in the nephrons for some time before being flushed into the bladder urine, the cells may
degenerate to a coarsely granular cast, later to a finely granular cast, and eventually, to a waxy
cast. Granular and waxy casts are believed to be delivered from renal tubular casts. The number
of casts is reported as “number and type seen per low power field (LPF)”. An example of a
report might read: “5-10 hyaline casts/LPF.”

Some epithelial cells from the skin surface or from the outer urethra can appear in the urine.
Some forms of crystals appear in the urine of healthy individuals. Abnormal crystals can indicate
liver disease or some forms of genetic abnormalities.

URINE DIPSTICK TEST

Reagant Storage and Stability

32
  Store at room temperature between 15-30℃ (59-86F)
 Do not use product after espiration date.
 Do not store the bottle in direct sunlight.
 All unused strips must remain in the original bottle.
 Do not mix lot number.
 Do not touch reagent areas of the reagent strips.

The reagent tests areas are ready to use upon removal from the bottle and the entire reagent strip
is disposable. The strips are to be read visually. Accurate timein g is essential to provide optimal
results. The reagent strip must be kept in the bottle with the cap tightly closed to maintain
reagent activity. To obtain optimal results, it is necessary to use FRESH, well-mixed,
uncentrifuged urine.

PROCEDURE:

1. Testing should be performed with adequate lighting. Inadequate light can lead to
incorrect color comparisons. Testing should not be done by those with limited color
discrimination, or those who have not yet been tested for color discrimination.
2. Collect FRESH urine specimen in a clean dry container. Label specimen container with
patient’s name preferably before giving it to the patient). Wear disposable medical
gloves.
3. Mix urine well immediately before testing.
4. Remove one strip from bottle and replace cap.
5. Completely immerse reagent areas of the strip in FRESH urine and remove immediately
to avoid dissolving out reagents.
6. While removing, run the edge of the strip against the rim of the urine container to remove
excess urine.
7. Hold the strip in a horizontal position to prevent possible mixing of chemicals from
adjacent reagent area and/or contamination.
8. Visually compare reagent area to corresponding Color Chart on the bottle label at the
time specified. HOLD STRIP CLOSE TO COLOR BLOCKS AND MATCH
CAREFULLY. Avoid laying srip directly on the Color Chart, as this will result in the
urine soiling the chart.

9. Proper read time is critical for optimal results.

 Glucose and bilirubin at 30 seconds after dippling.
 Ketone test at 40 seconds

33
  Specific gravity at 45 seconds
 pH, protein, urobilinogen, blood and nitrate 60 seconds
 Leukocytes at 2 minutes.

Results and Interpretation

Results with the Diagnostic Reagent Strips are obtained in clinically meaningful units directly
from the Color Chart comparison. All expected reference ranges are indicated on the Patient
Result Form

A routine urinalysis includes reporting of

1. Appearance: Clear, hazy, Cloudy, Turbid

2. Color: Colorless, Straw, Yellow, Amber, etc.
3. Specific Gravity: Note Result
4. Leukocyte Esterase: Negative, Trace, 1+, 2+, 3+
5. Nitrite: Positive or Negative
6. Ph: 5.0, 5.5, 6.0, .6.5, 7.0, 8.0
7. Protein: Negative, Trace, 1+, 2+, 3+, 4+
8. Glucose: Negative, Trace, 1+, 2+, 3+, 4+
9. Ketones: Negative, Trace, Small. Moderate Or Large.
10. Blood: Negative, Small, Moderate Or Large.
11. Urobillinogen: 0.2, 0.1, 2.0, 4.0, 8.0

Note: if results are not expected levels, send a specimen to the laboratory. Report to PDCT
coordinator any significant differences in POCT level vs. laboratory level.

Documentation:

1. All patient results are to be recorded on the Results Log and in the patient’s record.

34
 2. All reference ranges are noted on the Reference range sheet should be in each patient’s
chart.

Limitations:

As with all laboratory tests, definitive diagnostic or therapeutic decisions should be based on any
single result of method.

URINE PREGNANCY TEST:

The hCG pregnancy Test Strip is a test kit for the determination of hCG Human Chorionic
Gonadotropin) in urine specimens this test kit is used to obtain a visual qualitative result in the
early detection of pregnancy.

Human Choriotic Godaotropin is a glycoprotein hormone sereted by developing the placenta

shortly after fertilization. The appearance of hCG soon after conception and its subsequent rise in
concentration during early gestatonal growth make it an excellent marker for the detection of
pregnancy test.

TEST PROCEDURE:

1. To begin testing, open the sealed pouch by tearing along the notch. Remove the test
from the pouch. Note: First morning urine usually contains the highest concentration
of hCG and is therefore the best sample when performing the urine test. However,
random collected urine specimens may be used.
2. Holding the strip vertically, carefully dip it into the specimen (you may collect your
urine in a clean, dry container). Immerse the strip into the urine sample with the arrow
end pointing towards the urine. Do not immerse past the MAX Line (Market Line).
Take the strip out after 3 seconds and lay the strip flat on a clean, dry, non-absorbent
surface. (Note: in rare instances when dye does not enter the result are dip the tip of
the test strip in the urine as instructed above until the dye begins travelling across the
white result area.
3. Wait for colored bans to appear. Depending on the concentration of hCG in the test
specimen, positive results may be observed in as little as 40 seconds. However, to
confirm negative results, the complete reaction time of 5 minutes is required. It is
important that the background is clear before the result is read. Do not read results
after the specified reaction time.

INTERPRETETATION OF RESULTS

35
Negative: Only one color band appears on the control region. No Apparent band on the test
region. This indicates that no pregnancy has detected.

Positive: Distinct color bands appear on the control and test regions. Presence of both test line
and control line indicate that you are pregnant. The color intensity of the test bands may vary
since different.

RAPID HBS ANTIGEN STRIP TEST

36
INTENDED USE:

The Rapid Test HBsAg) in human resum as an aid in the diagnosis of hepatitis B infection. Test
results are read visually without any instrument. This test is intended primarily as an initial
screening test. Any specimen found to be repeatedly reactive should be confirmed by
neutralization procedures utilizing human antiHBs.

PRINCIPLE

This assay is a chromatographic immunoassay (CIA), containing filter membrane coated with
anti-HBs antibodies and colored gold colloidal reagents label with anti-HBs.

There are two regions, test region and control region, on the membrane of the test strip. The test
line (T), a purple color band in the test region of membrane, will be developed rapidly (form
30seconds to 10 minutes) when HBsAg is present in the specimen. If HBsAg is below 2 ng/mL
or not present, no T line will be developed in the test region. The control line (c), a purple color
band in the control region of the test, should always appear regardless of the presence of HBsAg.
Serving as an internal qualitative control of the test system.

RAPID HBS ANTIGEN STRIP TEST

37
REAGENTS AND MATERIALS PROVIDED:

 Test strips (Each strip of one single test)

STORAGE CONDITIONS

The test should be stored at refrigeneration or room temperature (8* to 30*C) in the sealed pouch
with a desiccant packet for the duration of the shelf life. Freezing or expose the kit to
temperature over 30℃ may cause malfunction.

PRECAUTIONS

1. For in vitro diagnostic use only.

2. Do not use test kit beyond expiration date.
3. Do not open the test strip foil ouch until you are ready to perform the test.
4. Icteric, lipemic, haemolysed, heat treated and contaminated sera may cause erroneous
results.

SPECIMEN COLLECTION

Serum and plasma heparin, citrate, or EDTA) specimens may be used.

HBsAg is thermo-labile. Specimens containing particulate matter or red blood cells may give
inconsistent results and should be centrifuged before testing (recommending 8,000-10,000 RCF*
x 10 minutes). Specimens which are not tested within 24 hours should be removed from the clot
or red blood cells.

BLOOD TYPING PROCEDURE

FORWARD TYPING:

38
 1. Mark a clean glass-slide as Anti A and Anti B
2. Place one drop of blood on each marked side of the side.
3. Add two drops of Anti-A serum to the blood on the side marked Anti-A
4. Add two drops of Anti-B serum to the blood on the side marked Anti-B.
5. Using an applicator stick, mix each side of the reaction.
6. Rock the slide back and forth and observed for agglutination.
7. Record the observation.

REVERSE TYPING:

 Make a clean glass slide as A cells and B cells.

 Place one drop of patient’s serum on each marked side of the side.
 Add one drop of A cells to the serum on the side of the side marked A cells.
 Add one drop of B cells to the serum on the side of the side marked B cells.
 Mix and examine for the presence of agglutination.
 Record the observation.

RH BLOOD TYPING:

1. Place two drops of blood on glass slide.

2. Add one drop of Anti-D serum. Mix and spread over the slide.
3. Place the glass directly on the glass place plate of a viewing box containing a light globe.
This will bring the temperature of the plate to 40-45C.
4. Rotate back and forth the slide and observed for agglutination.
5. Test is read and completed within 2 minutes.

STOOL Analysis

39
 A stool analysis is a series of tests done on a stool (feces) sample to help diagnose certain
conditions affecting the digestive tract. These conditions can include infection (such as from
parasites, viruses, or bacteria), poor nutrients absorption, or cancer.

For a stool analysis, a stool sample is collected in a clean container and then sent to the
laboratory. Laboratory analysis includes microscopic examination, chemical tests, and
microbiologic tests. The stool will may be examined for hidden (occult) blood, fat, meat fibers,
bile, white blood cells, and sugars called reducing substances. The pH of the stool also may be
measured. A stool culture is done to find out if bacteria may be causing an infection.

CLINICAL CHEMISTRY:

Control Storage and Stability

 Store at 2-8℃ before initial use. Do not freeze. When stored at 2-8℃, the control are
stable until the expiration date stated on the label.
 On initial use remove the controls from the refrigerator and allow to come to room
temperature (25-25℃), about 15-30 minutes
 After the initial use, the opened Control Bottles are to be stored at room temperature. Do
not store above 30℃. (86℉).
 When stored at room temperature (20-25℃) the controls are stable for one month. Room)
temperature expiration date must be noted on the control bottle label when the bottle is
opened.

Discard the controls and open a fresh vial if they are turbid or there is any evidence of microbial
contamination (discoloration, unusual odor) is present.

40