Sop-Clinical Pathology

R.K.Life Services Pvt.Ltd.
(Department of Laboratory Medicine)

Doc No. RKLS/SOP/02 SOP FOR CLINICAL PATHOLOGY
R.K. Life Services Pvt. Ltd

(Quality Manual for Department of Laboratory Medicine)
Meena Bhawan Kanchan Rd, Opp. Bora Service, G. S. Rd, Guwahati-781007
Telephone No: 2461474/73, Fax No: 2459601
e-mail : guwahati@theapolloclinic.com
STANDARD OPERATING
PROCEDURE
(CLINICAL PATHOLOGY)
Copy No: MASTER COPY

Holder: QUALITY MANAGER
Issue No: 04
Issue Date: 09.03.2020
Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

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Rev. No.: 00 Rev. Date: Nil
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TABLE OF CONTENT
LAST
SL REVISION
TITLE REVISION PAGE NO
NO NO
MADE
1 Urine Routine Examination 00 00 3–8
2 Stool Routine Examination 00 00 9 – 12
Sputum Routine
3 00 00 13 – 14
Examination
4 Cell Count Test 00 00 15 – 16
5 Bence Jones Protein 00 00 17 – 18
6 Semen Analysis 00 00 19 – 21
7 Total Protein in 24 hrs Urine 00 00 22
8 Quality Control Procedure 00 00 23
9 Safety and Precautions 00 00 23

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NAME OF TEST: URINE ROUTINE EXAMINATION

1. Purpose of examination
Urine Routine examination to include quantity, appearance, specific gravity, reaction,
albumin, sugar, bile salt, bile pigment, Urobilinogen, Occult blood and centrifuge
deposit for microscopy.
2. Principle of the Procedure used for Examination

a) Protein :
(i) Manual : Heat coagulation in acidic urine.
(ii)Dipstick: Indicator tetrabromophenol or tetrabromophenolphthalein blue show
coloured change from yellow, which is negative, to green- gold for a trace
reaction through green and then to greenish blue for elevated levels. to blue in
the presence of albumin.
b) Glucose :
(i)Manual: Benedicts reaction – When urine is boiled in alkaline copper
sulphate, glucose and other sugars reduce cupric ions to red colour cuprous
oxide.
(ii)Dipstick: First, glucose oxidase catalyzes the formation of gluconic acid and H 2O2
from the oxidation of glucose. A second enzyme, peroxidase, catalyzes the
reaction of H2O2 with a KI chromogen to oxidize the chromogen to colours
ranging from blue-green through green, brownish-green to brown.
c) Ketones : Strip and chemical test work on the same principle. Acetoacetate and
acetone react with sodium nitroprusside in an alkaline medium to give a violet
dye complex. The strips are more sensitive to acetoacetate than acetone.
d) Bilirubin : For Bile Pigment
(i)Manual: Barium chloride is used to precipitate the sulphates in the urine. Any
bilirubin present becomes attached to the precipitated barium sulphate. When
Fouchet’s reagent is added to the precipitate, the Ferric chloride oxydises the
Bilirubin to green blue coloured biliverdin.
(ii)Dipstick: Is based on the coupling of Bilirubin 2, 4 – dichlorobenzene –diazonium
fluroborate in an acidic medium. The colour ranges from tan to orangish when
no bilirubin is present through various shades of tan to reddish brown with
increasing levels of bilirubin.
Bilirubin : For Bile Salt
i) Bile Salt – add a pinch of sulphur power to urine in a test tube, if it sinks
record as positive and if it floats record as negative.
e) Urobilinogen :
(i)Manual:Urobilinogen reacts with p-dimethylaminobenzaldehyde to form a
red condensation product. The intensity of the colour produced
corresponds to the concentration of Urobilinogen present.
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(ii)Dipstick:The strip is specific for Urobilinogen and not affected by the

substances that interfere in the Ehrlichs test. A pink-red azody is
formed by 4-methoxibenzene-diazonium tetrafluoroborate when it
combines with Urobilinogen in a strong acid medium.
f) Hemoglobin :
(i)Dipstick:The test is based on the ability of hemoglobin to catalyze the
oxidation of colour indicator (tetramethylbenzidine) by an organic peroxide to
produce a colour ranging from greenish-yellow through bluish-green to dark
blue. Intact erythrocytes hemolysed on the test pad liberate hemoglobin and
produce blue-green dots on a yellow background.
g) Specific Gravity :
(i)Dipstick: The test pad contains two indicators, methyl red and bromothymol blue,
which give color changes ranging from blue-green to orange with increasing
ionic concentration. The strip specific gravity test does not indicate the amount
of non-ionic urinary constituents present such as urea, creatinine, or glucose.
h) pH :
(i)Dipstick: The test is based on the double indicators (methyl red and bromothymol blue)
which give colours ranging from red orange through green to blue overing the
urinary pH range of 5-9.
3. Performance specifications
Being a qualitative test these have not been worked out.
4. Primary Sample System

Urine: First morning sample is preferred, in case of urgency any mid-stream sample may
be tested. Urine should be collected in clean container. For Urobilinogen the best time to
collect the sample at 2 to 4 P.M. Urobilinogen reaches the highest concentration between 2
to 4 P.M.
5. Type of container & additives

Clean dry container without additives or detergent.
6. Required equipment and reagents

Glass Centrifuge Tube, Test Tubes, Slides, Cover Slips, Multistix, Benedict Reagent,
Acetic Acid, Sulphur powder, Fouchet’s Reagent, 10% H2SO4, 10% Barium Chloride.
7. Calibration procedure
Centrifuge and Urinometer is calibrated as per NABL norms.
8. Procedural Steps
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 The sample is received and properly labeled bearing name and ID number and test
to be performed.
 It is then transported to the Clinical Pathology Lab. The responsibility lies with the
reception. A test slip is generated and sent along with the specimen.
 Technician/Assistant perform the following test :-
 A number is assigned to each sample which also written on the test slip.
Colour :
The colour is recorded with following shades
(i) Colour less
(ii) Straw
(iii) Pale yellow
(iv) Yellow
(v) Deep orange
(vi) Red
(vii) Reddish
In the event the colour being different from the shades noted above, the Pathologist
should be consulted.
Appearance :
It is recorded as (i) Clear (ii) Turbid (iii) Smokey
Quantity :
The amount received is noted in ml. (an approximate figure is sufficient)
The sample is well mixed and 10ml labeled Centrifuge tube is filled with urine and it is
spun at 1800rpm for 3 minutes. The supernatant is poured into another similarly labeled
tube.
A dip stick (Multistix) is dipped into the tube containing the supernatant and the
following parameters are recorded :
i) Reaction – is noted as Acidic or Alkaline
ii) Albumin – is noted as nil, trace, +, ++, +++, ++++.
iii) Sugar – is recorded as nil, trace, +, ++, +++, ++++.
iv) Ketones/Acetone – is recorded nil, trace, +, ++, +++.
v) Bile Pigments – is recorded as absent or present.
vi) Urobilinogen – is recorded as nil, +, ++, +++.
vii) Occult Blood – is recorded as nil, trace, +, ++, +++.
Cross Checks :
(i) In case the urine is deep yellow and the bile pigment is negative then the following tests
is to be done.

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5ml of urine is added to 10ml of 10% Barium Chloride in a test tube. The precipitate is
collected in a filter paper by allowing the solution to pass through the filter paper. One or
two drops of Fouchet’s reagent is added to the filter paper, if green colour appears it is
recorded as positive.
(ii) If it tests positive for sugar Benedict’s tests is performed.

Add 8 drops of urine to 5 ml of benedicts solution. Hold the test tube against a flame and
allow it to boil. Cool the test tube and observe the change in colour, record as follows :
Blue,clear or cloudy Nil
Green, no precipitate Trace
Green, with precipitate (+)
Brown and cloudy (++)
Orange and cloudy (+++)
Red and cloudy (++++)
(iii) If it tests positive for albumin the following test to be done :
2/3rd of a test tube is filled with urine. Upper 1/3 rd of the filled urine is hold against a
flame, if turbidity appears add 1 to 2 drops of 10% Acetic Acid then heat again (Lower
1/3rd of the test tube acts as a control). If the turbidity still persists it is positive for
albumin and it is recorded as below :
Trace – Barely visible turbidity
(+) - Can see and read prints through the tube.
(++) - Can see prints but cannot be read.
(+++) - Cannot see print.
(++++) - Boiled solid.
Microscopic Examination :
 The deposit in the Centrifuge tube is placed on an appropriately labeled slide and
covered with a cover slip, the slides and the test slip is given to the pathologist for
microscopic examination.
 The slide is to be examined under 40X lens and record as below the following:
Red Blood Cell, Epithelial Cells, Pus Cells – These are recorded as number per high
power field as (a) Occasional – if less than 1/hpf, (b) In number/hpf and (c) Plenty – if
the number cannot be counted.
 Record presence or absence of crystals with type.
 Crystals are of following types. Only Cystine, Uric acid, Leucine and tyrosine crystals
are clinically significant.
 Casts is recorded in number/hpf with type.
 Yeast cells, bacteria, flagellates, spermatozoa etc. is recorded, if seen.
 The data is entered in Test Slip and send for typing.
9. Quality Control Procedure

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a) Liquicheck Urinanalysis control level 1 & 2 from BioRad is checked daily for internal
quality control.
b) Inter laboratory comparison is done with 2 NABL accredited laboratory once in 6
months.
10. Interferences
a) Protein :
(i) Dipstick: Dipstick test for protein may show false positive result if urine is
contaminated with disinfectants containing quarternary ammonium
compounds or chlorhexidine.
(ii) Strong alkaline urine may give false positive results.
(iii)Contamination with vaginal or urethral secretions may give false positive
results.
(iv) After infusion of plasma expanders like polyvinyl, pyrolidone.
b) Glucose :
(i) Oxygen receptors in urine like ascorbic acid and acetoacetic acid can give false
positive results.
(ii) Catalase will present in high concentration in the urine with E.coli infections and
can destroy H2O2 resulting in false negative results.
(iii) Disinfectants such as bleach can oxidize the chromogen directly giving false
positive reaction.
c) Ketones : High specific gravity or low pH urine may give false trace positive
reaction.
d) Bilirubin :
(i) If urine has very high level of ascorbic acid false negative result may be seen.
(ii) Beet root dye can interfere with this reaction. This dye becomes colourless in
alkaline urine.
(iii) Certain drugs can colour the urine deep orange resulting in interference.
e) Urobilinogen :
(i) Reaction with porphobilinogen is late.
(ii) With phenzaopyridine (found in some disinfectants) a rapid red colour is
found.
f) Hemoglobin : False positive reaction can result from the presence of contaminating
oxidizing detergents like bleach.
11. Principle of procedure for calculating result including Measurement uncertainty

None.
12. Biological Reference intervals

Specific Gravity – 1.010 – 1.030
Urobilinogen – Trace quantities are found in normal urine.

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13. Reportable interval of examination results

2 Hours at room temperature and 8 Hours at 4 – 80C.
14. Alert/Critical values, where applicable

Presence of Ketone in urine.
15. Laboratory interpretation

a) Infection
b) Jaundice
c) Hematuria
d) Presence of sugar
e) Ketosis
16. Safety precaution

General safety precautions as described in Quality System Procedure for sample
handling.
17. Potential source of variability

a) Contamination with vaginal or urethral secretions.
b) Dirty container
c) Drugs
d) Detergents in container
e) High level of Vitamin C.
REFRENCES:
i. Kaplan, L.A. and Pesce, A.J., Clinical Chemistry Theory Analysis and Correlation.
C.V.Mosby: 1004-1007; 1984.
ii. Brunzel, N.A., Fundamentals of Urine and Body Fluid Analysis. Philadelphia: Saunders;
1994.
iii. Graff, L.A., Handbook of Routine Urinalysis. Philadelphia: J. B .Lippincott Co.;1983.
iv. Jungreis, E., Spot Test Analysis. New York: John Willey & Sons; 1985.
v. Henry, J. B. et Al., Clinical Diagnosis and Management of Laboratory Methods, 18 th ed.
Philadelphia: Saunders; 1991.
vi. Scheer, W. D., American Journal of Clinical Pathology, 87: 86-93, 1987.
vii. Urinalysis and Collection, Transportation, and preservation of Urine Specimens; Approved
Guideline-Second Edition, GP 16-A2, Vol.21 No. 19, NNCCLS.
viii. Hepler, O.E., Manual of Clinical Laboratory Methods, 4th ed. Illinois: Thomas Books;

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NAME OF TEST : STOOL ROUTINE EXAMINATION
Physical, chemical and microscopic examination of the stool to include the following :
a) Colour
b) Consistency
c) Mucus
d) Blood
e) Reaction
f) Occult Blood
g) Reducing substances
h) Microscopic examination

a) Morphological identification of Ova and Parasite
b) For occult blood colour change in the test card.
c) Iodine colour change with starch
None.

Any random specimen of freshly voided stool may be examined and in case of occult
blood the patient is asked to be on vegetarian diet and stop Vitamin C for 3 days before
giving the sample.
If patient is constipated he is advised to take two tablets of dulcolax on the previous
evening. Sample should be collected in clean plastic container.

Clean dry container.

Glass slides, Test tubes, Microscope, Lugol’s Iodine, Saturated salt solution.
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None.
8. Procedural Steps
The sample is received at the reception desk. The container is checked and labeled with
name, ID number and test to be performed. The sample is then transferred to Clinical
Pathology Section for testing. The test procedure describe below is to be performed by
Assistant Technician.
a) Physical Examination –
The following parameters is to be tested:

(i) Colour – should be recorded in the following shades
Brown, yellow to yellow green, green, black, tan or cream, red. For any other shade seen
the pathologist must be consulted.
(ii) Consistency – is recorded as hard, soft, liquid, fatty or greasy. Under this heading if
following moieties if present is recorded (i) Gall Stone, (ii) Faecolith, (iii) Worms.
(iii) Frank Blood – Absence or presence of the same is noted.
(iv) Mucous – Absence or presence of the same is noted.
b) Chemical Examination :
pH and reaction – pH is tested by smearing a thin layer of stool on the pH paper or litmus
paper and read using comparative strips for pH and the colour change
from blue to red or red to blue of litmus paper is recorded. Red
Litmus Paper to Blue – Alkaline and Blue to Red is acidic. No change
is recorded as neutral.
Reducing Substances – 8 drops of liquid stool or equivalent is added to 5ml benedict’s
reagent and boiled and then cooled. Yellow green to orange colour
indicates positive results and is recorded as such. No change in colour
is recorded as negative.
TEST OF OCCULT BLOOD :
Range : It will detect approximately 10mg occult blood per 100ml. It is not specific for
hemoglobin.
Reagents : Hemospot: A Two filed test for occult blood in stool done by the standard
GUAIAC method.
Principle: If blood is present in stool the haematin in the hemoglobin molecule catalyses the
release of oxygen from the H2O2 which in turn oxidizes the colourless phenolic
components of gum GUAIAC to coloured quinones.During test, After the
addition of developer solution to the reactive surfaces of the result window, the
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reaction area turns blue if occult blood is present in the sample. If there is no
change in colour it indicates a negative result.
Procedure : Spread small amount of stool on the test card sample application window A &
B. Dry it. Turn the card, add developer solution to both windows and observe
colour change after two minutes.
Microscopic Examination of Stool -
Slide to be prepared, as described below and given to pathologist for examination.

Reagents required –
(i) Normal Saline (9gm/dl NaCl.)

(ii) Iodine (Dobell’s - Potassium Iodide 4g, Iodine 2g, distilled water 100ml.
Weigh the potassium iodide and dissolve in 50ml of distilled water, weigh the Iodine
and add to potassium iodide solution. Mix well to dissolve, and add the remaining water.
(iii) Saturated Solution of Sodium Chloride
Take a loop of normal saline and Iodine on properly labeled glass slide bearing the
number of the specimen. Add to it one loop full of representative stool specimen
containing mucous and blood if any. Take about 1g of stool in a 5ml container and add
saturated sodium chloride to it so as to fill the container. Place a cover slip on top of the
container and leave it for 10 minutes. Remove the cover slip and place it on a
appropriately labeled slide then send the slides and test slip to pathologist.
The microscopic examination is done by the pathologist and the following parameters are
recorded :
(i) Vegetable Cells

(ii) Starch Granules
(iii) Epithelial Cells
(iv) Pus Cells
(v) Red Blood Cells
(vi) Ova
(vii) Parasites
(viii) Cyst and Vegetative Forms
(ix) Fat Droplets
(x) Crystals
(xi) Undigested Muscle Fibre.
The data is entered in Test Slip and send for typing

a) Starch solution for checking Iodine and blood to check occult blood test reagent.
b) Inter laboratory testing once in two years.

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10. Interferences
None vegetarian diet and iron containing medicine can give false positive results.

None.

None.

8 Hours.

Occult blood test positive.

Microscopic examination.

handling.

Related to Occult blood test as described above and periodic shedding of Ova.
REFERENCES:
i. Knight et. al. (1989) Occult Blood Screening for colorectal cancer. JAMA, vol261, No.4, 587-
593.
ii. Rockey D et. al. (1998) Relative Frequency of Upper gastrointestinal and colonic lesions in
patients with positive occult blood tests. N Engl J Med 339; 153-159.
iii. Morris, W.D. et al (1976) Reliability of chemical tests for fecal occult blood in hospitalized
patients. Digestive diseases vol 21, No.10, 845-852.

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iv. Ostrow J. D. et al (1973) Sensitivity and reproducibility of chemical tests for fecal occult
blood with an emphasis on false positive reactions. Digestive diseases vol 18, No. 11, 932-939.

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NAME OF TEST : SPUTUM ROUTINE EXAMINATION
Microscopic and Macroscopic examination of Sputum.

Morphological identification of various components.
None.

Morning or random sample of sputum produced by deep coughing.

Clean dry preferably sterilized container.

Microscope, Slides, Gram Stain, ZN Stain, Gas flame and wire loop.
None.
8. Procedural Steps
The sample is received by the reception and labeled bearing the name, ID number and
name of the test. It is sent to clinical pathology lab. Following things are recorded.
a) Colour
b) Consistency
c) Odour
d) Presence or absence of the following is noted –
(i) Cheesy masses – Fragment of necrotic lung tissue.
(ii) Dittrichs Plugs – Are plugs from bronchi/bronchioles consisting of cellular
debris, fat and bacteria seen in bronchieactasis.
(iii) Curschmanns Spirals – These are twisted mucoid strands enclosing epithelial
cells, eosinophils and charcot leyden crystals.
(iv) Casts – Bronchial casts are molded in the bronchial and consist of fibrin and
mucin.
e) Makes smear of the sputum with wire loop on clean glass slides. Flame it to fix the
smears. Stain with the following stains :
(i) MGG/Lieshman
(ii) Gram Stain
(iii) Z.N. Stain
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Look under the oil emersion lens and record the following :
(i) Epithelial Cells
(ii) Inflammatory Cells
(iii) Red Blood Cells
(iv) Charcol Leyden Crystals – These are derived from eosinophils and are
hexagonal slender crystals with pointed ends.
(v) Elastic Fibres
(vi) Micro Organism (Bacteria)
(vii) Fungi
(viii) Acid Fast Bacilli
Record the above on test slip and send for typing.

a) All stains are controlled as described in the respective sections.
b) Interlab comparison is done once in two years.
10. Interferences
Contamination with saliva.

Not applicable.

Not applicable.

6 Hours.

None.

Not applicable.

handling.

Patient may not cough up the sputum and give only a superficial specimen.

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18. Reference: O.E Hepler. Manual of Clinical Laboratory Methods. Pg No 146, 147, 149

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NAME OF TEST : CELL COUNT OF FLUID
Estimation of total number of cells in (a) CSF (b) Peritoneal fluid (c) Pleural fluid and (d)
Synovial fluid

Neubaur’s Chamber count.
CV% 8.2 and range upto 100000/cmm.

The above samples to be collected in heparin vial at the ratio of 15±2.5 I.U. heparin/ml of
fluid. The collections is to be done by physician incharge.

Sterile container with heparin in the ratio 15±2.5 I.U. heparin/ml of fluid.

Neubaur’s counting chamber, Pasteur pipette and Microscope
None.
8. Procedural Steps
 Sample is checked for presence of coagulum before receiving. Coagulated samples

are not accepted.
 Neubaur Counting Chamber is charged with the fluid.
 1mm square is counted. Number of Cells x 10 = Number of Cells per cmm.
 Smear is made from Centrifuged deposit and stained with lieshman stain.
 Differential count is done on this smear.
 Result is recorded in test slip and sent for typing.

a) Clotted samples are not accepted.
b) Interlab testing, if possible.
10. Interferences
a) Clots
b) Hemolysis
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11. Principle of procedure for calculating result including Measurement uncertainty:
One big square of Neubar’s chamber measures 1 x 1mm and the depth is 0.1mm.

CSF Cell Count
a) Children and Adult – 0 – 6/cmm – All mono nuclear cells.
b) Infants - < 19/cmm
c) Neonates - < 30/cmm
Collection of fluid in other location occurs only in pathological condition.

8 Hours.

a) High Cell Count in CSF
b) High number of pus cells in any fluid

Not done.

handling.

a) Various disease condition
b) If sample is partially clotted
18. Reference: O.E Hepler. Manual of Clinical Laboratory Methods. Pg No 150, 151, 152.

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NAME OF TEST : BENCE JONES PROTEIN
Detection of Bence Jones Protein in urine.

The fact that Bence Jones Protein coagulates at 50 – 600C and dissolves at 900C.
None.

Urine sample, preferably first morning.

Any clean dry container.

Beakers, Test tubes, Thermometer, Tripod and Gas Burner.
Thermometer is calibrated.
8. Procedural Steps
 Urine is received and properly labeled and send to clinical pathology laboratory.
 Place a 250ml Beaker, half full with water on a Tripod and place a tube
containing the urine sample to be tested.
 Place a thermometer in the Beaker. Heat to raise the temperature to 50 to 600C.
 Look for turbidity in the urine at 50 to 600C.
 If turbidity appears continue heating till 90 to 100 0C when turbidity should
disappear.
 Allow the water to cool, turbidity will reappear on cooling to 600C.

Bence Jones Protein is positive with Sulphosalicylic Acid but negative with dipsticks.
10. Interferences
This test is relatively insensitive and requires minimum 150 mg/dl of Bence Jones
Protein.

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None.

None.

6 Hours.

Any positive results.

None.

handling.

It is not a very specific test.
18. Reference: O.E Hepler. Manual of Clinical Laboratory Methods. Pg No 8

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NAME OF TEST : SEMEN ANALYSIS
Analysis of Semen to include :
a) Volume
b) pH
c) Liquefaction time
d) Sperm count
e) Motility
f) Morphology

a) Counts are done with formal fixed spermatozoa
Not possible.

Sample collected by Masturbation after 3 days of sexual abstinence without any condom
in a sterile wide mouthed container. Note the time on the container label.
If it is collected at home it should be transported within 15 minutes.

Clean dry wide mouthed container.

Counting chamber, Microscope, Stains, diluting fluid etc.
a) Sperm count reagent

Sodium Bi-carbonate – 5g
40% Formalin – 1ml
Distilled water – 100 ml
b) Pap Stain
Pipettes are calibrated as per NABL norms.
8. Procedural Steps
 The patient is instructed on the method of collection when he comes for

appointment. Samples are not to be accepted if the collection is not done properly.

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 The patient is requested to give the sample within 10 minutes of collection or

collected in the laboratory itself.
 Before asking the patient to collect the sample ensure that the technician and
pathologist are present.
 The sample is labeled bearing patient’s name and ID number and test to be done and
immediately send to clinical pathology lab.
The technician to carry out the following tests :
(i) Makes a note of the collection time.

(ii) Volume is measure by drawing up the liquefied specimen in a pipette.
(iii) pH is measured using a pH paper.
(iv) The sperm count is done as follows :
a) Add 20 μl of liquefied semen to 380 μl of diluting fluid and let it stand for 5
to 10 minutes till all the mucous dissolves.
b) Charge the diluted sample in the Neubaurs counting chamber.
c) Count one large square (1 x 1mm).
Count x 200,000 = Count per ml
(v) Place one drop of liquefied semen on a slide and place a cover slip on it. Look
under the microscope under high power objective and record the percentage of
motile spermatozoa. Repeat the observation after four hours. Also record the
type of motility with their respective percentages as fast, slow and non-
progressive.
Make a smear of the semen and stain it with PAP Stain, look under the microscope and
report the morphology under following categories - (i) Normal (ii) Bent Head (iii)
Tapered Head (iv) Mid Piece Defect (v) Amorphous.
Data is recorded in the test slip and send for typing.

a) Test done by two persons and compared.
b) Interlab testing of morphology slides.
c) Abnormal semen results to be subjected to retesting on another occasion.
10. Interferences
a) Improper collection
b) Part of the specimen lost during collection
c) Condom collection

Neubar’s Chamber.

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 Volume – 2 – 3ml.
 pH – 6.4 – 8
 Count – 20 x 106/ml or more
 At least 50% should be of normal morphology
 More than 50% motile after 1 hour

Test to begin within 30 minutes.

None.

None.

handling.

a) Improper and incomplete collection
b) Delay in transport.
18. Reference: O.E Hepler. Manual of Clinical Laboratory Methods. Pg No 160

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ESTIMATION OF TOTAL PROTEIN IN 24 HOURS URINE
Purpose :
Total protein in 24 hours urine.
Performance Specification :
Total protein in 24 hours urine by Esbach’s method.
Primary Sample :
24 hours urine is used , preserved with Thymol crystal.
Container & Additives Requirement :

5 litre container with thymol crystals provided from the Apollo Clinic. Patient is asked
to store each voided urine after emptying the bladder at 7:00 AM to 7:00 AM of the
following morning.
Biological Referance Interval:

Normal(30-130)mg/24 hours
Equipment & Reagent :

Esbach’s reagent, Esbach’s Tube, Pippette, Filter paper.
Enviromental Condition :
At room temp.
Analytical Procedure & Result :

The urine is filtered until clear then fill Esbach’s tube with urine upto mark U and add
Esbach’s reagent upto mark R. The tube is closed with a rubber stopper and inverted 10
times slowly. Place in a test tube rack and keep in a cool place for sedimentation. Read at 24
hours. The readings ( R ) on the tube indicates grams of albumin per litre of urine.
Calculation: Total Protein/24hours = R x Volume of urine/1000
References :
Prepared By: No.
Issued By:
Approved By:
Hepler, O.E., Manual of Clinical Laboratory Methods, 4th ed. Illinois: Thomas Books
Section --A
QUALITY CONTROL PROCEDURE
A. Regular use of QC material of renowned QC kit manufacturer:

Daily internal QC: Biorad liqui check urinanalysis control is run daily (Level 1 & Level 2)
and as and when required.
B. Correlations of results:
Correlation is done with different parameters, which is documented.
Section --B
SAFETY AND PRECAUTIONS
 Safety instructions of different instruments and chemicals given by manufacturers

are followed strictly.
 General safety precautions of biohazard are observed.
 Samples are handled with utmost care.
 Samples are handled wearing gloves, facemask and apron.
 Reagents and chemicals are kept according to the company instructions

Prepared By: No.
Issued By:
Approved By:

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R.K.Life Services Pvt.Ltd.

(Department of Laboratory Medicine)

R.K. Life Services Pvt. Ltd

Copy No: MASTER COPY

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

1 Urine Routine Examination 00 00 3–8

2 Stool Routine Examination 00 00 9 – 12

4 Cell Count Test 00 00 15 – 16

5 Bence Jones Protein 00 00 17 – 18

7 Total Protein in 24 hrs Urine 00 00 22

8 Quality Control Procedure 00 00 23

9 Safety and Precautions 00 00 23

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

NAME OF TEST: URINE ROUTINE EXAMINATION

2. Principle of the Procedure used for Examination

(ii)Dipstick:The strip is specific for Urobilinogen and not affected by the

4. Primary Sample System

5. Type of container & additives

6. Required equipment and reagents

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

(ii) If it tests positive for sugar Benedict’s tests is performed.

9. Quality Control Procedure

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

11. Principle of procedure for calculating result including Measurement uncertainty

12. Biological Reference intervals

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

13. Reportable interval of examination results

14. Alert/Critical values, where applicable

15. Laboratory interpretation

16. Safety precaution

17. Potential source of variability

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

NAME OF TEST : STOOL ROUTINE EXAMINATION

2. Principle of the Procedure used for Examination

4. Primary Sample System

5. Type of container & additives

6. Required equipment and reagents

The following parameters is to be tested:

TEST OF OCCULT BLOOD :

Microscopic Examination of Stool -

Slide to be prepared, as described below and given to pathologist for examination.

(i) Normal Saline (9gm/dl NaCl.)

(i) Vegetable Cells

9. Quality Control Procedure

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

11. Principle of procedure for calculating result including Measurement uncertainty

12. Biological Reference intervals

13. Reportable interval of examination results

14. Alert/Critical values, where applicable

15. Laboratory interpretation

16. Safety precaution

17. Potential source of variability

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

Issue No. 04 Issue Date: 09.03.2020 Copy Page 7 of 23

NAME OF TEST : SPUTUM ROUTINE EXAMINATION

2. Principle of the Procedure used for Examination

4. Primary Sample System

5. Type of container & additives

6. Required equipment and reagents