STANDARD OPERATING PROCEDURE (SOP)

Legacy Pathology Laboratory, Kigali Rwanda

LEGACY DIRECT EXAMINATION OF STOOL FOR PARASITE

INFECTION SOP
CLINICS
PATHOLOGY Effective date: Pages: 8
LABORATORY LEG/PATH LAB/MIC-SOP-13-VERS …/…./….
001

Review date:

…../…../…….

Prepared by: Authorised by

Verified by:
Names: Name:
Title: …………………
Names :………… Laboratory Director
Signature :……………
Title::………………. Signature ………………….

Date:……………
Date :…………………
Date:……………..

REVISION HISTORY
Revised by Version Review date Effective date Description of change

NA 001 NA

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 STANDARD OPERATING PROCEDURE (SOP)

Legacy Pathology Laboratory, Kigali Rwanda

1. INTRODUCTION

Legacy Pathology Laboratory aims to adhere to the International Standards related to Good Laboratory
Practice and the ISO 15189 accreditation process guidelines. The Laboratory uses Standard Operating
Procedures (SOPs) to facilitate adherence to these regulations.

2. ABBREVIATIONS AND DEFINITIONS

LEG/PATHLAB: Legacy clinics pathology laboratory
QC: Quality control
SOP: Standard Operating Procedure
PAR:Parasitology
NaCl: Sodium Chloride
Wet preparation: Direct examination of a sample without any modification

3. USERS
Only Laboratory staff are authorized to use this procedure upon being trained and competently
ensured. However, the trainees should be allowed to use this SOP and the equipment under supervision.
4. PURPOSE OF THE EXAMINATION

Many parasites cause disease in human. Some of these parasites are excreted in stool; they are called
intestinal parasites. Intestinal parasites can be identified by examination of fresh stool samples. In
stool samples we can find worms (eg. Ascaris lumbricoides) and segments of worms (e.g. Taenia spp)
is visible to the eye. By microscopic examination of fresh stool samples, we can find eggs (e.g.
Hookworm) and larvae of worms (e.g. Strongyloides stercoralis). We also find protozoa trophozoites
(e.g. Amoeba) and cysts (e.g. Cyclospora cayetanensis). In heavy and moderate infection, a direct
smear examination with normal saline (0.9%) and/or iodine to stain cysts, is usually sufficient. For
light infections, a concentration of the stool sample might be required to find helminth (worm) eggs and
protozoa by microscopic examination.
5. PRINCIPLE AND METHODS OF THE PROCEDURE USED FOR EXAMINATION
When stool is emulsified in physiological saline (NaCl 0.9% w/v), parasites are conserved and
separated from fecal debris. Therefore, they can be easily detected, identified and counted using a light
microscope.

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 STANDARD OPERATING PROCEDURE (SOP)

Legacy Pathology Laboratory, Kigali Rwanda

6. PERFORMANCE CHARACTERISTICS
(NA)
7. TYPE OF SAMPLES
Fresh stool sample at least 3g collected into a suitable size, clean, dry, leak-proof container.

8. PATIENT PREPARATION
( NA)
9. TYPE OF CONTAINER AND ADDITIVES
Stool container with applictor is required

10. REQUIRED EQUIPMENT AND REAGENTS

- Glass slides

- Cover slip (20 x 20 mm)

- Wooden applicator

- Grease pencil.

- Microscope.

- Normal Saline (0.9% NaCl and iodine solution)

11. ENVIRONMENT AND SAFETY CONTROL

Standard precautions for handling infectious agents should be observed when performing this test. All
samples must be considered as hazardous materials. Their handling needs all safety precautions,
especially the use of appropriate personal protective equipment (e.g. gloves, lab coat, etc…).
12. CALIBRATION PROCEDURES (METROLOGICAL TRACEABILITY )
( NA)
13. PROCEDURAL STEPS
a. Wear appropriate PPE

b. Gather necessary materials and reagents

c. Record the sample in the laboratory register

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 STANDARD OPERATING PROCEDURE (SOP)

Legacy Pathology Laboratory, Kigali Rwanda

d. Record the macroscopic appearance (presence of blood or mucus, texture (hard, soft, watery), smell,
color)

e. In case of several samples, immediately begin to analyze those that are liquid or show presence of
mucus or blood

f. Prepare and label a slide for each sample

g. If stool is watery, place a drop of stool directly on the slide, cover and examine on the microscope.

h. If stool is hard or soft, place a drop of NaCl 0.9% or iodine solution 1% on slide

i. Using a spatula, remove a small portion of sample (about the size of a match head) and mix with the
drop of saline or iodine and cover.

j. Examine preparation under the 10x and 40x objectives

k. Identify any parasites found (mature, eggs, cysts, etc) or any other abnormalities

l. Record and report the result to the appropriate service

m. Discard wastes and wash hands

14. QUALITY CONTROL PROCEDURES

Use positive slides and diagrams to compare

15. INTERFERENCES
(NA)
16. PRINCIPLE OF PROCEDURES FOR CALCULATING RESULTS
Count and report the number of parasites per High Power Field, Low Power Field or per Preparation
17. BIOLOGICAL REFERENCES INTERVALS
(NA)

18. REPORTABLE INTERVAL OF EXAMINATION RESULTS

(NA)
19. INSTRUCTIONS FOR DETERMINING QUANTITATIVE RESULTS WHEN A RESULTS
IS NOT WITHIN THE MEASUREMENT INTERVAL

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 STANDARD OPERATING PROCEDURE (SOP)

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(NA)

20. ALERT/CRITICAL VALUES, WHERE APPLICABLE

(NA)
21. LABORATORY CLINICAL INTERPRETATION

Many pathogenic parasites are excreted in stool. Often, when a person is infected with intestinal
parasites, other symptom such as anaemia, eosinophilia, diarrhoea and malabsorption are also
present. However diagnosis by physical examination is not sufficient to identify intestinal parasitic
infection. Stool examination is essential to identify parasites that cause the disease.

22. CLINICAL INTERPRETATION

 For negative preparation, report: Negative

 For Positive preparation, the report is done as follow:

Motile trophozoites may be identified to the species level.

 Eg: Giardia lamblia trophozoites .

 Protozoan cysts may be identified to the species level,

 Eg: Entamoeba Coli cysts .

Helminth eggs may be identified.

 Eg: Ascaris lumbricoides eggs .

23. POTENTIAL SOURCES OF VARIATION

- Old Specimen or Sample delay; e.g. Amoebic trophozoites begin to degenerate within 1-2 hours
after collection. Always examine watery and blood-stained specimens first.

- High Temperature: Cysts, flagelates and eggs also undergo changes especially if the stool is left at
high temperatures; ambient temperature is sufficient.

- Specimens contaminated with dirt, water or urine. Urine or water may be damaging to trophozoites.

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 STANDARD OPERATING PROCEDURE (SOP)

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- Never use tincture iodine as it contains alcohol and it would destroy amoeba trophozoites

- Wrong identification: Refer to pictures if you are in doubt about structures that resemble eggs,
cysts or trophozoites.

24. REFERENCES

- WHO (1991) Basic Laboratory Methods in Medical Parasitology. WHO, Geneva

- Chessbrough, Monica (1998). Laboratory Practice in Tropical Countries. Tropical Health
Technology, Cambridgeshire.
- Sherchand, Jeevan B. et.al. (1998) A cross Sectional Study on Intestinal Parasitic
Infections in Primary School Children of Godar VDC Nepal. 1-1-98, p.61-64.
JONAMELS, Kathmandu.

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 STANDARD OPERATING PROCEDURE (SOP)

Legacy Pathology Laboratory, Kigali Rwanda

26.SIGNING PAGE

We, staff members of LEG/PATHLAB, do hereby certify that we have read, discussed and
understood the content of this SOP. We commit ourselves to abide by its spirit and shall strive to
comply and make it complied with.

N° Staff Names Staff Signature Date

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17

I, …………………………………, being Laboratory Director , do hereby certify that all the

staff, as listed above, have read, discussed and understood the content of this SOP as indicated
herein.

Signature & Stamp: Date: / /2021

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 STANDARD OPERATING PROCEDURE (SOP)

Legacy Pathology Laboratory, Kigali Rwanda

27.AMENDMENT PAGE

Date Amendment Number Amendment Page Responsible

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