Dengue IgG/IgM Card Test

R For Professional Use

A Rapid Card Test to detect IgM & IgG antibodies to Dengue Virus in human Serum or Plasma or Whole blood
IVD
Read the pack Insert before use provided along with the kit
REF BDC

INTENDED USE : Dengue IgG/IgM rapid card test is a solid phase CONTENTS OF THE KIT :
immunochromatographic assay for the rapid, qualitative and Pack Size 5 Test 10 Test 20 Test 50 Test
differential detection of IgG and IgM antibodies to dengue virus in 1. Test Device 5 Nos. 10 Nos. 20 Nos. 50 Nos.
human serum or plasma or whole blood. This test is intended for 2. Assay Diluent 0.75 ml 1.5 ml 3.0 ml 7.5 ml
Professional use as aid in the presumptive diagnosis between 3. Silicagel 5 Nos. 10 Nos. 20 Nos. 50 Nos.
5µl Dropper
primary and secondary dengue infection. This test provides only a 4. 5 Nos. 10 Nos. 20 Nos. 50 Nos.
5. Pack Insert
preliminary test result. Therefore, isolation of virus, antigen 1 No. 1 No. 1 No. 1 No.

detection in fixed tissues, RT-PCR and serological test like MATERIAL REQUIRED BUT NOT PROVIDED:
haemagglutlnation-Inhibition test, more specific alternative a) Sterilized vial b) Disposable gloves
diagnosis method must be used in order to obtain a confirmation of c) Micro pipette(1-10ìl) d) Sodium hypochlorite solution (free
dengue virus Infection. This kit can detect all 4 Dengue serotypes available chlorine 50-500mg/L) e) Autoclaved Tips
by using a mixture of recombinant dengue envelope proteins.
WARNINGS & PRECAUTIONS :
INTRODUCTION : The Dengue Virus belongs to the Flavivirus In order to obtain reproducible result , the following rules must be
group of viruses transmitted by the mosquito, Aedes aegypti and observed :
Aedes albopictus mosquitoes, widely distributed throughout the a). Read this Pack insert carefully.
tropical and subtropical areas of the world. The dengue fever is very b). DO NOT FREEZE THE KITS. If refrigerated the kits should be
common in rainy season. The symptoms of Dengue fever are brought to room temperature before testing . Assay should be
sudden onset of fever, headaches, pain in the back and limbs, conducted between 15 - 30ºC.
lymphaderopathy, maculopapular rash and retrobulbar pain. There c). Do not use umbilical cord blood because it prevents colloidal
are four known distinct serotypes (dengue virus 1,2, 3 and 4). In gold from migrating & can interfere with result.
children infection is often subclinical or causes a self-limited febrile d). Do not use the kits beyond their expiry date.
disease. However, if the patient is infected a second time with a e). Use whole blood.
different serotype a more severe disease, dengue hemorrhagic f) Carefully serve the prescribed number of drops to be added,
fever or dengue shock syndrome is more likely to occur. Dengue is 5ìl of serum or plasma or whole blood and 2 drops of diluent only.
considered to be the most important arthropod-borne viral disease g). Use the test device soon after it is removed from the pouch.
due to the human morbidity and mortality it causes. Traditionally h).Do not use the test device, if the pouch seal is broken.
haemagglutination-inhibition (HAI) test has been the most I). Avoid any contamination among samples; for this purpose,
commonly used serological assay for dengue diagnosis. disposable tips and sterilized vial should be used for each sample
and reagent.
Primary Dengue infection is associated with mild to high fever,
j). Read the result in 20 minutes.
headache, muscle pain and skin rash. Immune response includes
IgM antibodies produced by 5th day of symptoms and persist for k). Do not smoke, eat drink or apply cosmetics during the assay.
30-60 Days. IgGs appear in the 14th day and persist for life. l) For in vitro Diagnostic Use only.
Secondary infections often result in high fever and in many cases m). For single use only.
with hemorrhagic events and circulatory failure. Secondary n).Do not use the whole blood sample, which is stored for more
infections show that IgGs rise within 1-2 days after the onset of than 3 days as it may give false positive result.
symptoms and induce IgM after 20 days of infection. Traditionally,
HAI titers have been used to classify infection. The current All human serum and plasma samples should be considered
definitions as primary or secondary depends on an assay of paired potentially infectious. It is recommended that all specimens of human origin
serum specimens separated in time by at least 7 days, although should be handled as recommended for any potentially infectious
any acute specimen with a HAI titer>1:2560 is defined as coming human serum or blood specimen in the Centers for Disease
from a patient with secondary Flavavirus infection. Control / National Institute of Health manual “Biosafety in
Microbiological and Biomedical Laboratories”,1984. Never
TEST PRINCIPLES : The Dengue IgG/IgM test kit is a sandwich pipette by mouth. Do not smoke, eat or drink in areas in which
immunochromalographic membrane based screening test to specimens or kit reagents are handled. Afterwards wash hands
detect the antibodies for dengue virus. The test can be used with carefully. Avoid splashing or forming aerosols. Discard all
whole blood. The test employees the use of colloidal gold particle materials and specimens as if capable of transmitting infection
conjugated with IgG & IgM binding proteins and a unique The preferred method of disposal is autoclaving for a minimum of
combination of dengue antigen immobilized on the membrane at T. one hour at 121°C.
After the addition of sample and running buffer in the sample well of Liquid wasted not containing acid may be mixed with sodium
test device, the mixture passes through the conjugate gold and hypochlorite so that the final mixture contains 50-500 mg/I
makes a immune complex. available chlorine. Allow 30 minutes for decontamination. .
The Immune complex will bind with the antigen immobilized at test Note : Liquid waste containing acid must be neutralized with a
zone(T) of the test device if antibodies to dengue present in the proportional amount of base prior to the addition of sodium
sample. The remaining complex continues to migrate to a control hypochlorite. Spills should be wiped up throughly using either an
area (C) in the test device and produces a colored band In the iodophor disinfectant or sodium hypochlorite solution. Materials
control (C) zone. The control band indicates that the test has been used to wipe up spills should be added to biohazardous waste
performed properly and is in working condition. Appearance of matter for proper disposal. Reagents are stored between 2-30°C.
pink/purple band at test (T) in addition to band at control (C) Avoid unnecessary exposure to light. Do not use reagents after
indicates that sample is positive for dengue antibodies. expiration date. Do not mix or interchange reagents from different
kits or kit batches. Cross contamination of reagents or samples
STORAGE AND STABILITY : Kit should be stored between 2°C- can cause erroneous results.
30°C in the sealed pouch. The test kit is stable until the expiration Use a new pipette tip for each sample. Optimal results are
date mentioned on the pouch when stored under these conditions. obtained by strictly adhering to the test protocol. Accurate and
Do not use the test device, if the pouch is damaged or the seal is precise pipetting, as well as following the exact time and
broken. Keep the test device in the sealed pouch until ready for use. temperature requirements is essential.
The opened diluent bottle is stable for 2 years.
 Once the assay has been started, all steps should be performed EXPECTED VALUE : Primary dengue is characterized by the
without interruption. presence of detectable IgM 3-5 days after the onset of
SPECIMEN : Fresh serum or plasma or whole blood infection. Secondary dengue is characterized by the elevation
Specimen collection & Preparations (Collection by Veni Puncture) : of specific IgG 1- 2 days after the onset of infection and in the
a) Collect whole blood into a sterilized vial (Containing EDTA, majority of cases this is generally accompanied by an elevation
Citrate or Heparin) by venipuncture. of IgM.
b) If specimens are not immediately tested, they should be
refrigerated at 2-8°C. Sensitivity: 98.4%
c) When stored at 2-8° C the whole blood sample should be used Specificity: 99%.
with in 3 days.
LIMITATIONS OF THE TEST
COLLECTION USING A LANCET : 1. This test detects the presence of antibodies to dengue in the
a) Clean the area to be lanced with an alcohol swab. Specimen and should not be used as the sole criteria for the
b) Squeeze the end of the finger and pierce with a sterile lancet. diagnosis of Dengue virus infection.
c) Wipe away the first drop of blood with sterile gauze or cotton. 2. In early infections and some secondary infections, detectable
d) Using the micropipette or dropper draw 5ìl of whole blood. levels of IgM antibodies may be low. Some patients may not
produce detectable levels of antibody within the first 7-10 days
ASSAY PROCEDURE : after infection. Where symptoms persist, patients should be
Remove as many test units from the pouches as needed and put on retested 3-4 days after the first specimen.
flat surface. Label them with sample/patient identity. 3. If the test result is negative and clinical symptoms persists,
Add 5ìl of serum or plasma or whole blood to the sample window of additional follow-up testing using other clinical methods is
the device. recommended. A negative result does not preclude the possibility
Add 2 drops of the Diluent provided in the dropper bottle by of an early infection of Dengue virus.
holding the bottle vertically over the sample well. REFERENCES :
Read the results in 20 minutes only. 1. Songee L. ranch and Paul N. Levett. Evaluation of four
methods for detection of immunoglobulin M antibodies to dengue
virus. Clin. Diagn. Lab. Immunol. Vol6 (4) p 555-557, 1999.
2.Lamm S.K. (1995), dengue haemorhagic fever. Rev. Med.
Micro, 6-39-48.
IgG IgM c 3. Seth, J. (1991. standardization & quality assurance. In
Dengue

Principle and practice of immunoassay, Ed. C.P. Price & D.J.

Newman. Macmillan Publishers, pp.154-189.
4. Dengue haemorrhagic fever : Diagnosis, treatment, prevention
and control. WHO 2nd Edition 1997.
INTERPRETATION OF TEST RESULT :
NEGATIVE: Only Red band appears in the control zone NOTE:: Even after the best effort is made to supply the product as
(C) of the result window. No band at test zone (T). per the sample submitted but due to continuous R & D, the
company reserves the right to improve/change any
IgG IgM c specifications/components without prior information/notice to the
Dengue

buyer
LIMITED EXPRESSED WARRANTY OF MANUFACTURER
The manufacturer limits the warranty to this test kit, as much as that
the test kit will function as an in vitro diagnostic assay within the
Nature of Sample, Procedure limitations and specifications as
POSITIVE : Appearance of two colored bands, one in the test zone described in the product instruction manual, when used strictly in
(IgM and or IgG) and another in the control zone (C), indicates that accordance with the instructions contained. The manufacturer
sample is positive for antibodies of dengue virus. disclaims any warranty expressed or implied including such
expressed or implied warranty with respect to merchantability,
Positive for IgG & IgM Positive for IgM fitness for use or implied utility for any purpose. The manufacturer’s
liability is limited to either replacement of the product or refund of
Dengue

Dengue

IgG IgM c IgG IgM c

the purchase price of the product and in no case liable to 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
Positive for IgG any injury, damage or economic loss, howsoever caused by the
product in the use or in the application there of.
Dengue

IgG IgM c
BS ISO-15223-1:2012(E) MEDICAL DEVICES SYMBOL
30°C
Temperature Date of In vitro Diagnostic
IVD
Limitation Manufacture Device
2°C 2001-06

Company name Consult Instructions

LOT Batch Code
INVALID : If a colored band does not appears in the control zone or & address For Use

Only Test Line appears, then the result is Considered Invalid. Use by Company Name EC REP
Authorised
Representative in
Company European Community
R-12, 2013-05-03

Sufficient for KEEP AWAY FROM

Dengue

Dengue

IgG IgM c IgG IgM c 2 Do Not Reuse

SUNLIGHT

KEEP DRY NON-STERILE NEGATIVE

NON CONTROL
STERILE CONTROL

POSITIVE
CONTROL CONTROL

®
Manufactured in India by :

11-A, 4th Cross, Veerasandra Industrial Area, Electronics City,

Bangalore - 560100, Karnataka, INDIA Tel.: +9180 3319 4000 (30 lines)
Fax : +9180 3319 4001 www.bhatbiotech.com