One Step Fecal Occult Blood Test
WARNINGS & PRECAUTION
Cat. No:
1) In vitro diagnostic use for professional use only.
2) Do not use test kit beyond the expiry date.
3) The test device should not be reused.
4) Patient specimens may contain infectious agents and
should be handled as though capable of transmitting
disease. Wear disposable gloves throughout the specimen
collection and assay procedures.
INTENDED USE
The One Step Cassette Style Fecal Occult Blood Test is a simple,
direct binding immunoassay for the rapid and qualitative detection
of human occult blood in feces. The sensitivity is very high with the
ability to detect 50 ng/mL of human hemoglobin in feces. In
addition, the accuracy of the test is not affected by interfering
substances, and dietary restriction is not necessary. For
professional laboratory use only.
SUMMARY AND EXPLAINATION
The test principle is an immunochromatographic sandwich method,
which employ two specific monoclonal antibodies to selectively
identify hemoglobin in test samples. The result is very specific, and
easier to interpret than those of guaiac-based test.
The assay is conducted by adding diluted stool specimen to the
test device and observing the formation of colored lines. The
specimen migrates via capillary action along the membrane to
react with the colored conjugate. Positive specimens react with the
specific colored antibody conjugates and form a colored line at the
test line region of the membrane. Absence of this colored line
suggests a negative result. To serve as a procedural control, a
colored line will always appear at the control line region if the test
has been performed properly.
REAGENTS
1)
2)
3)
Test reagent
One Step Fecal Occult Blood Test
Sample Collection Device with buffer
Instructions for use
C
T
SPECIMEN COLLECTION
Collect stool sample by using the special sample collection
device provided. First, unscrew the top of the sample collection
device, take out the sample collection stick, and collect the
sample by dipping the stick into 3 different places of the stool
sample. Then, put the sample collection stick back in the
sample collection device and screw together tightly.
INTERPRETATION OF RESULTS

N Negative: Only one colored band appears on the control
(C) region. No apparent band on the test (T) region. This
means that FOB level is below 50ng/mL.

Positive: In addition to a pink colored control (C) band, a
distinct pink colored band will also appear in the test (T)
region. This indicates that the FOB concentration is more
than 50ng/mL. Please consult your physician to perform a
much more detailed examination.

Invalid: None of line appears or no line appears in the
control (C) region. An invalid result may be due to improper
testing procedures or deterioration of the kit components.
Repeat the assay sequence using a new device.
If the sample cannot be tested on the day of collection, store the
stool sample at 4°C. Stir and bring the specimen to room
temperature before testing.
DIRECTIONS FOR USE
1.
2.
3.
4.
Control region: Goat anti-mouse (IgG) polyclonal antibody
Test region: mouse monoclonal anti-hemoglobin antibody A
5.
Colloidal gold conjugate of monoclonal anti-hemoglobin antibody B
squeeze
Stool sample
MATERIALS PROVIDED
Coated Antibodies:
Labeled Antibody:
break off
the tip
shake
Break point
The assay is designed to detect lower levels of colorectal bleeding
than other biochemical detection methods.
PRINCIPLE OF TEST
Dip stool sample
6.
Remove the test device from its foil pouch by tearing
along the notch and use it as soon as possible.
Specimen
collection.
See
also
SPECIMEN
COLLECTION
Shake the sample collection device several times.
Holding the sample collection device upright, carefully
break off the tip of collection device.
Squeeze 3-4 drops (~100ul) of the sample solution on
the test sample pad, as in the illustration.
Read the test results after 5-10 minutes. It is important
that the background is clear before the result is read. Do
not read results after more than 20 minutes.
C
T
Negative
C
T
C
T
Positive
C
T
Invalid
STORAGE AND STABILITY
The test kit can be stored at temperatures between 2 to 30C
in the sealed pouch to the date of expiration. The test kit
should be kept away from direct sunlight, moisture and heat.
The expiration dating was established under these storage
conditions.
Revision date: March 2006
QUALITY CONTROL
A colored line appearing in the control region (C) is the
internal procedural control. It confirms sufficient specimen
volume and correct procedural technique. A clear background
is an internal negative background control. If the test is
working properly, the background in the result area should be
white to light pink and not interfere with the ability to read the
test result.
External controls may also be used to assure that the
reagents are working properly and that the assay procedure
is followed correctly. It is recommended that a control be
tested at regular intervals as good laboratory testing process.
Users should follow the appropriate federal, state, and local
guidelines concerning the running of external quality controls.
LIMITATIONS
1. As with all diagnostic tests, all results must be considered
with other clinical information available to the physician. A
definite clinical diagnosis should only be made by the
physician after all clinical and laboratory findings have
been evaluated. This test cannot intended to replace G.I.
fibroscope, endoscopy, colonoscopy, or x-ray analysis.
2. This test is limited to the detection of Fecal Occult Blood in
human stool sample only.
3. Although the test is highly accurate in detecting human
hemoglobin, a low incidence of false positive results may
occur. In addition, because many bowl lesions, indulging
some polyps and colorectal cancers, may bleed
intermittently or not at all, occult blood may not be
uniformly distributed throughout a fecal sample. Thus rest
results may be negative even when disease is present.
PERFORMANCE CHARACTERISTICS
Accuracy
This test is sensitive for 50 ng/mL human hemoglobin in stool
sample. 100 spiked stool samples with concentrations at 0 ng,
25 ng, 50 ng, 75 ng, 100 ng / ml were tested. The results in
Table 1 showed that the cutoff was the expected 50 ng/mL.
Moreover, two types of abnormal blood (Thalassemia and
Sickle Cell) were tested and no false negative issues were
raised as results were as expected.
One Step FOB Test kit for relative sensitivity and specificity in
648 stool samples. Only 2 samples were discordant, the
agreement is 99.67%. (Table 1)
Table 1: Cutoff Concentration for IND FOB Test
Hemoglobin
(ng/mL)
# of Samples
Negative
Postive
0
25
50
75
100
20
20
0
20
20
0
20
2
18
20
0
20
20
0
20
Interfering Substances
A performance study was completed to investigate the cross
reactivity of other species of hemoglobin and tissue extracts
on the FOB Test. The hemoglobin species of bovine, equine,
pig, rabbit, sheep, fish, chicken, and goat origin was added to
the test device and the results as expected. Hemoglobin of
the species was added to normal stool extracts at both 0 and
50 ng/ml human hemoglobin and the results were negative
and positive respectively. In addition, the study was repeated
with tissue extracts of beef, pig, rabbit, sheep, fish, chicken
and goat and no cross reactivity was evident. Lastly, toilet
water with the presence of various cleaners (enzymatic to
chlorax based) was added to test device and results were as
expected following the same protocol as above.
A performance study was completed to investigate the
interference of dietary substances on the FOB test. Aqueous
extracts of raw broccoli, cauliflower, cantaloupe, horseradish,
red radish, parsnip, and turnip were added to test device.
Also included were dietary iron, vitamin C and horseradish
peroxidase. The dietary substance extracts were added to
normal stool extracts at both 0 and 50 ng/ml human
hemoglobin and the results were as expected. All of the 0
ng/ml Hb samples spiked with interfering substances were
negative, while all of the 50ng/ml Hb spiked samples were
positive.
Table 1: Comparison between Syntron Bioresearch FOB
and IND FOB Test
IND kits
Syntron
kits
Subtotal
+
28
2
30
+
-
0
618
618
Subtot
al
28
620
648
References:
Gorzynski TJ, Krco CJ, David CS., An idiotope expressed on
a monoclonal antibody specific for human haemoglobin beta
chain and naturally occurring immunoglobulin(s). J
Immunogenet. 1985 Dec;12 (6):281-91.
Gorzynski TJ, Krco CJ, David CS., The Hb-2d cross-reactive
idiotype. A common idiotype expressed by monoclonal
and polyclonal antibodies to human haemoglobin.
J Immunogenet. 1985 Dec;12(6):267-79.
GRAPHICAL SYMBOLS USED
Storage temperature
Lot number
In vitro diagnostic device
Expiry date
Read instruction before use
Manufacturer
D
Do not reuse
Method Comparison
To establish the sensitivity and specificity of IND Diagnostic
One Step FOB Test kit relative to other rates of qualitative
FOB tests, 648 clinic samples were studied. Another
commercially available qualitative test kit (Syntron
Bioresearch FOB) was used to compare with IND Diagnostic
Revision date: March 2006