HEPACARD
One Step Rapid Visual Test For the Qualitative Detection of HBsAg in Human Serum / Plasma
INTENDED USE protected from exposure to humidity.
HEPACARD is a visual, rapid, sensitive and accurate one step
WARNING FOR USERS
immunoassay for the qualitative detection of Hepatitis B Surface
CAUTION: ALL THE SAMPLES TO BE TESTED SHOULD BE HANDLED
Antigen (HBsAg) in Human Serum or Plasma. The assay is intended to
AS THOUGH CAPABLE OF TRANSMITTING INFECTION. NO TEST
be used as an aid in the recognition and diagnosis of acute infections
METHOD
and chronic infectious carriers of the Hepatitis B Virus (HBV).
INTRODUCTION
The antigenic determinant of the HBsAg protein moiety is antigenically
heterogenous and it determines specific HBV serotypes and provides a
basis for immunodetection. The principal antigenic determinant is “a”
which is common to all HBV serotypes. In addition, two pairs of subspecific
determinants have been identified, d/y & w/r, which are apparently mutually
exclusive. Four antigenic combinations are therefore possible: adw, adr,
ayw and ayr.
PRINCIPLE
HEPACARD is a one step immunoassay based on the antigen capture, or
“sandwich” principle. The method uses Anti-HBsAg antibodies
conjugated to colloidal gold and Anti-HBsAg antibodies immobilized on a
nitrocellulose strip in a thin line. The test sample is introduced to and
flows laterally through an absorbent pad where it mixes with the signal
reagent. If the sample contains HBsAg, the colloidal gold-antibody
conjugate binds to the antigen, forming an antigen-antibody-colloidal gold
complex. The complex then migrates through the nitrocellulose strip by
capillary action. When the complex meets the line of immobilized
antibody (Test line) “T”, the complex is trapped forming an antibody-
antigen-antibody collidal gold complex. This forms a pink band indicating
the sample is reactive for HBsAg. When the complex meets the line of the
corresponding immobilized monoclonal antibody on test lines, the complex
is trapped forming a purplish pink band which confirms a reactive test result.
Absence of a coloured band in the test region indicates a non-reactive
test result. A red procedural control line should always develop at ‘C’
region to indicate that the test has been performed properly.
MATERIAL PROVIDED
1. Hepacard (individually pouched) 2. Sample Dropper
3. Instruction Manual
KIT PRESENTATION
20 Test Pack 100 Test Pack
DESCRIPTION OF SYMBOLS USED
The following are graphical symbols used in or found on J. Mitra
diagnostic products and packing. These symbols are the most common
ones appearing on medical devices and their packing. They are
explained in more detail in the European Standard EN ISO 15223-
1:2016.
Manufactured By In vitro
diagnostic
medical device
No. of tests See Instruction for use
Lot Number Temperature
Batch Number Limitation
Manufacturing Date Caution, see
instruction for use
Expiry Date Catalogue Number
Do not use if package Keep away from
sunlight is damaged
Single use only Keep Dry
STORAGE AND SHELF LIFE
HEPACARD should be stored at 2-30ºC in the cool and driest area
available. Expiry date on the kit indicates the date beyond which the kit
should not be used. The kit should not be frozen & must be
CAN OFFER COMPLETE ASSURANCE THAT HUMAN BLOOD
PRODUCTS WILL NOT TRANSMIT INFECTION.
1. The use of disposable gloves and proper biohazardous clothing is
STRONGLY RECOMMENDED while running the test.
2. In case there is a cut or wound in hand, DO NOT PERFORM THE
TEST.
3. Do not smoke, drink or eat in areas where specimens or kit
reagents are being handled.
4. Tests are for in vitro diagnostic use only and should be run by
competent person only.
5. Do not pipette by mouth.
6. All materials used in the assay and samples should be
decontaminated in 5% sodium hypochlorite solution for 30-60
min. before disposal or by autoclaving at 121ºC at 15psi for
60 min. Do not autoclave materials or solution containing
sodium hypochlorite. They should be disposed off in
accordance with established safety procedures.
7. Wash hands thoroughly with soap or any suitable detergent,
after the use of the kit. Consult a physician immediately in
case of accident or contact with eyes, in the event that
contaminated material are ingested or come in contact with
skin puncture or wounds.
8. Spills should be decontaminated promptly with Sodium
Hypochlorite or any other suitable disinfectant.
9. Do not open the foil pouch to remove the product until it
attains room temperature and you are ready to perform the
test. In case of five test pouch packing, seal the pouch
containing balance devices with the help of clamp and rod
provided, every time they are opened.
10. Take out the Cards from the pouch just before performing
the test to avoid denaturation of antisera due to atmospheric
exposure.
Optimal test performance requires strict adherence to the
test procedure described in the insert.
SAMPLE / SPECIMEN COLLECTION & STORAGE
a) HEPACARD should be performed on human serum or plasma only
immediately after collection.
b) If not tested immediately, specimen should be refrigerated at 2-8 oC
upto 3 days following collection.
c) If testing within 3 days is not possible, specimen should be
stored frozen at -20oC.
d) Specimen containing visible precipitates or cloudy specimens
may give inconsistent test results. Such specimens should be
cleared prior to testing by high speed centrifugation i.e. 10,000
rpm for 15 minutes before testing.
e) Haemolysed specimen or specimen with microbial
contamination should be discarded and fresh aliquot should
be collected.
TEST PROCEDURE R.T.
1. Bring the required number of HEPACARD foil pouches 20-30o
and specimen to room temperature prior to testing.
2. Take out HEPACARD device from the foil pouch.
3. Label the test card with patient’s name or
identification number.
4. Add 2 drops (70 µl) of human serum/plasma
specimen into the sample well using the
dropper provided (use separate
dropper/microtip for each specimen).
5. Allow reaction to occur during the next 20 minutes.
6. Read results at 20 minutes.
7. Discard the HEPACARD immediately after reading result at 20 (B) EXTERNAL EVALUATION
minutes, considering it to be potentially infectious. External evaluations were carried out at two different centres and the
INTERPRETATION OF RESULT results are as mentioned below:
REACTIVE : i) M/s PATH (Program for Appropriate Technology in Health),
As shown in Fig.1, appearance of pink coloured Seattle, USA. Sensitivity : 100%
line, one each in test region “T” and control Specificity : 100%
region “C” indicates that the sample is ii) Christian Medical College, Vellore, India (a reference centre for Govt. of
REACTIVE India)
Fig. 1
for HBsAg. A difference of intensity in colour Sensitivity : 100%
may occur between the Test line & Control line depending on the Specificity : 100%
concentration of the HBsAg in the serum but this does not affect This information is provided for the Scientific Community Enquiring for
interpretation of the result. an independent evaluation other than company’s in-house evaluation. It is
Depending on the concentration of HBsAg, positive results may be not for commercial or promotional purpose.
observed within 60 seconds. However, to detect concentration around (C) PRECISION
0.5 ng to 1ng/ml Within-run (Intra assay) & between-run (Interassay) precisions have
and to confirm a negative result, the test result should be read been determined by testing 15 replicates of four samples: 5 negative
only at 20 minutes. If the conc. of HBsAg in the sample is very high, and 10 HBsAg positive samples; 3 weak, 6 medium and 1 strong
only test line may be observed. This is due to Hook’s effect. Such positive. The C.V.(%) of negative, weak, medium and strong positive
samples should be diluted 1:10 or 1:20 in normal saline & again re-run the samples were within 10% of time.
test, Diluted sample should show both control & test line. In case, if control
line does not appear or is faint dilute the sample further. In case of ACKNOWLEDGMENT
faint test line (’T’), centrifuging the sample at 10,000 rpm. J. MITRA & CO. PVT. LTD. WISHES TO ACKNOWLEDGE THAT HEPACARD
for 15 minutes and repeat the test using fresh card. Consider a HAS BEEN DEVELOPED UNDER TECHNOLOGY TRANSFER AGREEMENT
faint test line also as positive result. WITH M/s PATH
(Program for Appropriate Technology in Health) SEATTLE U.S.A.
NON-REACTIVE :
As shown in Fig.2 appearance of one Reserve Bank of India approval for this technology transfer has been
distinct pink line in the control region “C” granted vide Regn. No: DFT 97 NDR 0012.
only, indicates that the sample is “NON
REACTIVE” for HBsAg.
Fig. 2
INVALID : LIMITED EXPRESSED WARRANTY DISCLAIMER
When neither control line nor the test The manufacturer limits the warranty to the test kit, as much as that
line appears on the membrane as the test kit will function as an in vitro diagnostic assay within the
shown in Fig.3, the test should be treated limitations and specifications as described in the product instruction-
as invalid which may be because of Fig. 3 manual, when used strictly in accordance with the instructions
following reasons: contained therein. The manufacturer disclaims
a) Improper storage at temperature other than the recommended No. of Status Hepacard Hepacard
temperature. Samples (+ ve) (- ve)
b) Wrong test procedure. 225 All ELISA 225 -
c) Long atmospheric exposure of the test device after opening the +ve
pouch.
3240 ALL ELISA - 20 3220
d) Turbid or lipaemic sample. ve
The test should be repeated using a new HEPACARD after
centrifugation of test sample. SENSITIVITY: 100% SPECIFICITY: 99.38%
LIMITATIONS OF THE PROCEDURE
1. The HEPACARD is for in vitro diagnostic use only.
2. The test should be used for the detection of HBsAg in serum or
plasma only and not in other body fluids.
3. This is only a Screening test. All reactive samples should be
confirmed by confirmatory test. Therefore for a definitive
diagnosis, the patient’s clinical history, symptomatology as well as
serological data, should be considered. The results should be
reported only after complying with above procedure.
4. Additional follow up testing using available clinical methods (along with
repeat HEPACARD test) is required, if HEPACARD test is non-
reactive with persisting clinical symptoms.
5. False positive results can be obtained due to the presence of Rf
antibodies, patients with auto-immune disease, liver problems, renal
disorders and antenatal samples.
PERFORMANCE CHARACTERISTICS
(A) INHOUSE EVALUATION
(i) Analytical Sensitivity: The sensitivity of Hepacard is 0.5 ng/ml
(ii) The performance of Hepacard has been evaluated in house with
fresh as well as frozen HBsAg negative and positive samples. The
testing has been done with clinical samples, samples from
random blood donors, cross reacting samples; RA, CRP, ASO,
antenatal and patients with diseases related to HBV. The results
of in-house studies are as follows:
any warranty expressed or implied including such expressed or
implied warranty with respect to merchantability, fitness for use or
implied utility for any purpose. The manufacturer’s liability is
limited to either replacement of the product or refund of the
purchase price of the product and in no case liable to for claim
of any kind for an amount greater than the purchase price of the
goods in respect of which damages are likely to be claimed. The
manufacturer shall not be liable to the purchaser or third parties
for any injury, damage or economic loss, howsoever caused by
the product in the use or in the application there of.
BIBLIOGRAPHY
1. Blumberg, B.S., (1964) Bull. N.Y. Acab Med., 40:377
2. Blumberg B.S. etal, (1965) J.A.M.A. 191:541.
3. Caldwell C.W. etal., (1977) Clin. Chem. Acta: 31:305.
4. Peterson, D.L. etal., (1982) J. Biol. Chem., 257(17): 10414.
5. Robin, E (1979) Fed. Proc. 33 (13) 2665.
R-10
VER-
in vitro diagnostic Reagent, not for medicinal use
01
J. Mitra & Co. Pvt. Ltd.
Rev. Date: Oct.-
MN/HPC/011
A-180-181, Okhla Ind. Area, Ph.-1, New Delhi-110
020, INDIA Ph.: +91-11-47130300, 47130500
e-mail: jmitra@jmitra.co.in Internet: www.jmitra.co.in
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