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SOP-002 Procedure
1.0 PURPOSE
This Standard Operating Procedure (SOP) provides guidelines for performing macroscopic and
microscopic examination of stool specimens to detect fecal occult blood, aid in the detection of early
stages of gastrointestinal problems, determine the cause of anemia, and screen for early stages of
colon cancer.
2.0 SCOPE AND APPLICATION
This procedure involves two main components of fecalysis:
● Macroscopic examination of stool characteristics
● Microscopic examination using normal saline solution
● Chemical examination for occult blood and other parameters
This procedure is applied for routine fecalysis testing of all patient stool specimens requiring analysis.
3.0 REFERENCES
Davidsohn, MD, Israel, Clinical Diagnosis by Laboratory Methods, 15th Edition, W.B. Saunders Co.,
Philadelphia, pp.18-45
Kaur, K., Zubair, M., & Adamski, J. J. (2023, April 23). Fecal occult blood test. StatPearls - NCBI
Bookshelf.
4.0 DEFINITION OF TERMS
Occult Blood Blood in stool that is not visible to the naked eye
pH A measure of the acidity or alkalinity of urine
Consistency Physical form of the stool (formed, hard, soft, or watery)
Mucus A thick, slimy substance that may indicate inflammation
Urobilinogen A product of bilirubin metabolism found in stool
Porphyrins Compounds involved in the synthesis of heme
Trypsin A digestive enzyme that breaks down proteins
Bilirubin A product of red blood cell breakdown
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Enzyme Technique A cross-match technique that uses enzymes (papain or cysteine) to
enhance agglutination.
Occult Blood Presence of blood in urine that may not be visible to the naked eye
RBC/HPF Red blood cells per high power field
WBC/HPF White blood cells per high power field.
5.0 RESPONSIBILITIES
Medical Technologist/ MT/ To macroscopic and microscopic examination of stool specimens
Lab Technician according to established procedures
6.0 MATERIALS AND EQUIPMENT
Equipment
● Microscope
● Timer
Specimen
Type of Specimen: Fresh stool
● Volume of specimen: thumb/ peanut size (3-5 grams) stored in clean/ sterile, dry, non-
breakable, sealable and leak proof wide mouth screw cap container
Reagents
● Normal Saline Solution
● Positive and Negative Stool Controls
Glasswares
● Glass slides.
Miscellaneous
● Gloves
● Fecal occult blood test kits
● Glass slides
● Blowipes or paper towel
● Marker/pen
● Applicator stick
● Clean/sterile containers with wide mouth screw cap
7.0 PROCEDURE
Macroscopic and microscopic analysis of stool specimen is a rapid and convenient method of
detecting fecal occult blood aids in the detection of early stages of gastrointestinal problems,
determination of the cause of anemia and to screen for early stages of colon cancer.
Refer to SOP-015- Receipt and Performance of Laboratory Tests
7.1 Specimen Collection
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7.1.1. Collect fresh stool specimen (3-5 grams, thumb/peanut size)
7.1.2. Place in clean, sterile, wide-mouthed container
7.1.3. Label with patient's full name, date, and time of collection
7.1.4. Ensure specimen is uncontaminated with:
- Urine
- Tissue paper
- Toilet water
- Other body secretions
7.2 Macroscopic Examination
7.2.1. Observe and record:
- Color (brown, green, black, or red)
- Consistency (formed, hard, soft, or watery)
- Presence of mucus
- Amount
- Odor
- Size and shape
- Presence of gross blood, pus, or parasites
7.3 Microscopic Examination
7.3.1. Place 1 drop of normal saline on slide
7.3.2. Add small portion of stool specimen
7.3.3. Mix with applicator stick and spread on slide
7.3.4. Cover with coverslip
7.3.5. Examine under microscope for:
- Fat/neutral fat/crystals and soaps
- Undigested food
- Meat fibers
- Starch
- Eggs and segments of parasites
- Yeasts
- Leukocytes
8.0 TEST ANALYSIS
8.1 Guaiac-Based Methods
I. Positive Result: Blue color appears within 30-60 seconds in test area
II. Negative Result: No blue color appears in test area
III. Invalid Result: No color in positive control area or unexpected color in negative control area
8.3 Result Interpretation Time Frames
I. Results must be read within 30-60 seconds
II. Blue color begins to fade after 2-4 minutes
III. Do not interpret results after 4 minutes
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9.0 REFERENCE VALUES
9.1. Macroscopic Parameters
- Amount: 100-200 g/day
- Color: Brown
- Consistency: Plastic, formed
- Gross blood, mucus, pus, parasites: None
9.2. Microscopic Parameters
- Fat: 18% fatty acid (colorless)
- Undigested food: None to small amount
- Meat fibers, starch, trypsin: None
- Parasites: None
- Yeasts: None
- Leukocytes: None
Component Normal Range
Water Up to 75%
pH 6.5-7.5
Occult Blood Negative
Urobilinogen 50-300 mg/24hr
Porphyrins:
Coporphyrins 400-1200mg/24hr
Uroporphyrins 10-40 mg/hr
Nitrogen <2.5g/24 hr
Bile Negative in adults; positive in children
Trypsin 20-250 mOsm
Electrolytes
Sodium 5.8-8 mEq/24 hr 2.5-3.9 mEq/24 hr 15.7-20.7 mEq/24
Chloride hr
Potassium
Lipids 0-6 g/24 hr
Refer to SOP-014-Reporting of Results for the next process in reporting these results based from
the normal values.
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10.0 PRECAUTIONS
10.1. Perform regular calibration and maintenance of equipment used
Refer to SOP-010-Program for the Calibration, Preventive Maintenanace, and Repair of
Instruments
10.2. Run quality control samples to ensure accuracy of test results.
Refer to SOP-016-Reporting of Workload and Quality Control
10.3. Results must be read within 30-60 seconds
10.4. Blue color begins to fade after 2-4 minutes
11.0 LIMITATIONS AND INTERFERENCES
11.1 Specimen Limitations
I. Specimens must be fresh
II. Avoid contamination with urine or other body secretions
III. Samples should be collected before contact with toilet water
IV. Multiple samples (3 different days) recommended due to intermittent bleeding
V. Dried samples can be stored at room temperature for up to 14 days
VI. Moist samples may show false-negative results due to heme degradation
11.2 Dietary Interferences
I. Raw fruits and vegetables containing peroxidases (turnips, broccoli, horseradish, cauliflower,
cantaloupe, parsnip, red radish)
II. Red meat consumption (3-day restriction recommended)
III. Vitamin C supplements (>250mg/day) may cause false-negative results
IV. Cooking vegetables at 100°C for 20 minutes inactivates plant peroxidase activity
11.3 Medication Interferences
I. Acetylsalicylic acid
II. Unfractionated or low-molecular-weight heparin
III. Warfarin
IV. Clopidogrel
V. Nonsteroidal anti-inflammatory drugs
VI. Selective serotonin reuptake inhibitors
VII. Iron supplements
VIII. Corticosteroids
IX. Cancer chemotherapeutic agents
11.4 Environmental Interferences
I. Toilet sanitizers may cause false-positive results
II. Chlorine-generating sanitizers affect guaiac methods
III. Povidone-iodine antiseptic solutions cause false-positive results
IV. Exposure to extreme heat or humidity
V. Prolonged contact with toilet water
11.5 Patient Conditions to Consider
I. Active hemorrhoids
II. Menstruation
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III. Urinary tract infection
IV. Hematuria
V. Peptic ulcer disease
VI. Altered bowel movements
11.6 Method-Specific Limitations
I. Guaiac-based methods are less sensitive for upper GI bleeding
II. Immunochemical methods may not detect upper GI bleeding due to globin degradation
III. Heme-porphyrin testing is better for detecting upper GI bleeding
12.0. APPROVAL
Prepared: Reviewed:
Angelie Leonado, RMT Nice Jane Noynay, RMT
CHIEF MEDICAL TECHNOLOGIST
Approved:
Kayen Tampus, RMT, MD
PATHOLOGIST
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