ST.
CECILIA DIAGNOSTIC CLINIC
FECAL OCCULT BLOOD TEST
(OnSite FOB-HI Rapid Test)
1.0 INTRODUCTION
The objective of this procedure is to check stool samples for hidden (occult) blood. It aids in
the detection of bleeding caused by a number of gastrointestinal disorders.
2.0 TEST PRINCIPLE
The OnSite FOB-HI Rapid Test is an iFOBT designed to specifically detect low levels of
human fecal occult blood. It is a lateral flow chromatographic immunoassay for the qualitative
detection. The test strip in the cassette consists of:
1. A colored conjugate pad containing monoclonal anti-hHb antibody
conjugated with colloid gold (anti Hhb conjugates)
2. A nitrocellulose membrane strip containing a test line (T line) and a control
line (C line)
The T line is pre-coated with another monoclonal anti-hHb antibody, and
the C line is pre-coated with a control line antibody.
When adequate volume of test specimen is dispensed into the sample well of the test cassette,
the specimen migrates by capillary action across the cassette. hHb, If present in the specimen at
or higher than 25 ng/mL, will bind to the anti-hHb conjugates. The immunocomplex is then
captured by the pre-coated reagent forming a colored T line, indicating a FOB POSITIVE test
result.
Absence of the T line suggests a NEGATIVE result. Each test contains an internal control (C line)
which should exhibit a colored line of the immunocomplex of the control line antibodies
regardless of the color development on the T line. If the C line does not develop, the test result
is INVALID, and the specimen must be retested with another device.
3.0 SAFETY AND ENVIRONMENTAL PRECAUTIONS
All human blood and other potentially infectious material (OPIM) are to be treated as
infectious and handled according to “ standard precautions”. Therefore, all personnel handling
specimens or other materials should wear appropriate Personal Protective Equipment’s (PPEs)
which include but not limited to: Gloves, Gowns, Masks
4.0 SPECIMEN
PATIENT PREPARATION
1. Specimens should not be collected from patients with the following conditions
which may interfere with the test results:
*Menstrual bleeding
*Bleeding hemorrhoids
*Constipating bleeding
*Urinary bleeding
2. Dietary restrictions are not necessary.
3. Alcohol and certain medications such as aspirin, indomethacin, phenylbutazone,
reserpine, corticosteroids and nonsteroidal anti-inflammatory drugs may cause
gastrointestinal irritation and subsequent bleeding and produce POSITIVE reactions.
On the advice of a physician, these medicines may be temporarily discontinued for
7 days prior to and during the test period.
COLLECTION AND HANDLING
- Consider any materials or human origin as infectious and handle them using
standard biosafety procedures.
- When receiving patient’s Fecal Occult Blood Test request, instruct the patient
to:
1. Collect a random sample of feces in a clean, dry receptacle.
2. Fill in all required information on the patient ID sticker and apply to
the stool collection device.
-Label clean screw top container with:
*Full Name
*Date of Birth
*Date and Time of collection
- Instruct the patient to do handwashing before and after collection
and make sure that the specimen doesn’t touch the inside of the
toilet.
3. Open the stool collection device by unscrewing the top and use the
collection stick to randomly pierce the stool specimen in at least five
different sites. Do not scoop stool specimen. Ensure that stool
specimen is only in the grooves of the collection stick. Excess stool
specimen may lead to invalid test result.
4. Replace the collection stick and tighten securely to close the stool
collection device.
5. Shake the stool collection device vigorously to extract the hHb in the
specimen.
NOTE: It is recommended to test the specimen immediately after
extraction. If not tested immediately, the extracted specimen may be
stored at room temperature (20-37 degree Celsius) for up to 10 days
or at 2-8 degree Celsius for up to 21 days. For longer storage, the
extracted specimen may be frozen at -20 degree Celsius. Avoid
multiple freeze-thaw cycles.
5.0 EQUIPMENTS AND MATERIALS
5.1 Individually sealed foil pouches containing:
a. One cassette test device
b. One desiccant
5.2 Stool collection devices, each containing 2 mL of sample extraction
buffer (REF SB-R2011)
5.3 Patient ID sticker
5.4 Instructions for use
MATERIALS MAY BE REQUIRED AND NOT PROVIDED
Positivia FOB Rapid Test Control Kit (Cat # C2011) contains positive
control and negative control.
MATERIALS REQUIRED BUT NOT PROVIDED
Clock or Timer
A container to hold fecal specimen
6.0 PROCEDURE
6.1 Bring the specimen and test components to room temperature if refrigerated or
frozen.
6.2 When ready to test, open the pouch at the notch and remove device. Place the test
device on a clean, flat surface.
6.3 Shake the stool collection device vigorously to ensure a homogenous liquid
suspension
6.4 Hold the stool collection device vertically. Twist off the tip. Dispense 2 drops (70-
90UuL) of the solution into the sample well of the cassette. Do not overload samples.
6.5 Set up timer
6.6 Results can be read at 10 minutes. POSITIVE results can be visible in as short as 1
minute. NEGATIVE results must be confirmed at the end of the 10 minutes only.
However, any results interpreted outside 10 minutes should be considered INVALID
and must be repeated. Discard used device after interpreting the result following local
laws governing the disposal of device.
7.0 QUALITY CONTROL
7.1 Internal Control
- This test contains a built-in control feature, the C line. The C line develops after
adding sample. If the C line does not develop, review the whole procedure and
repeat test with a new device.
7.2 External Control
- Good Laboratory Practice recommends using external controls, positive and
negative, to assure the proper performance of the assay, particularly under the
Following circumstances:
a. A new operator uses the test kit, prior to performing testing of
specimens
b. A new lot of test kits is used.
c. A new shipment of test kits is used
d. The temperature during storage of the kit falls outside of 2-30 degree
Celsius.
e. The temperature of the test area falls outside of 15-30 degree Celsius
f. To verify a higher than expected frequency of positive or negative
results
g. To investigate the cause of repeated invalid results
8.0 INTERPRETATION OF ASSAY RESULTS
1. NEGATIVE RESULT
- If only the C line develops, the test indicates that the concentration of hHb in
the sample is below 25 ng/mL in buffer. The result is NEGATIVE or NON-
REACTIVE.
2. POSITIVE RESULT
- In addition to the presence of the C line, if the T line develops, the test
indicates that the concentration of hHb in the sample is equal to or higher
than 25 ng/mL in buffer. The result is FOB-HI POSITIVE or REACTIVE
Specimens with POSITIVE results should be confirmed with alternative testing
method(s) and clinical findings before a diagnosis is made
3. INVALID
- If no C line develops, the assay is INVALID regardless of any color
development on the T line. Repeat the assay with a new device. If caused by
an excess amount of fecal specimen collected, collect a new specimen and
retest
9.0 LIMITATIONS OF THE TEST
The Test Procedure and the Interpretation of Assay Result sections must be
followed closely with testing for the presence of occult blood in feces. Failure
to follow the procedure may give inaccurate results
The OnSite FOB-HI Rapid Test is to aid in diagnosis and is not intended to
replace other diagnostic procedure such as G.I fibroscope, endoscopy,
colonoscopy or X-ray analysis. Test results should not be deemed conclusive
with respect to the presence or absence of gastrointestinal bleeding or
pathology. A POSITIVE result should be followed up with additional
diagnostic procedures to determine the exact cause and source for the occult
blood in the feces
A NEGATIVE or NON-REACTIVE result can be obtained even when a
gastrointestinal disorder is present. For example, some polyps and colorectal
cancers may bleed intermittently or not at all during certain stages of the
disease. A NEGATIVE or NON-REACTIVE result can also be obtained if the
quantity of occult blood present in the specimen is below the detection limit
of the assay
The OnSite FOB-HI Rapid Test has not been validated for testing of patients
with hemoglobinopathies
Specimens containing visible blood may produce NEGATIVE results due to the
hook effect
The results obtained with this test should only be interpreted in conjunction
with other diagnostic procedures and clinical findings
10.0 REFERENCES
1. America Cancer Society, Inc. Cancer Reference Information: Can Colon and Rectum
Cancer be Found Early? (online) Available: http://www.cancer.org
2. Allison JB, Takawa IS, Ransom LJ, Adrian AL. A comparison of fecal noccult blood
tests for colorectal-cancer screening. N. Eng. J. Med. 1996; 334:155-159.
3. Saito H. Screening for colorectal cancer by immunochemical fecal occult blood
testing (Review). Jpn J. Cancer Res 1996; 87:1011-1024
ST. CECILIA DIAGNOSTIC
CLINIC
OnSite FOB-HI Rapid Test
( Instruction for use)
Horacio A. Saguil, Jr., MD Lourdes Anicia Cardema-Soller, MD
Pathologist/ Head of Laboratory Medical Director/Owner
Prepared by:
TITLE
Janine Donna Karan
Melmida, RMT
Reviewed / Approved by:
Horacio Saguil, MD SECTION:
Head Pathologist CLINICAL MICROSCOPY
Lourder Cardema-Soller, MD STANDARD OPERATING
Medical Director PROCEDURE
Effectivity Date: April 04, 2022 ST. CECILIA DIAGNOSTIC CLINIC
FECAL OCCULT BLOOD TEST
Revision No.:
0
Number of Pages:
6
