Dengue NS1/IgG/IgM Test (GICA) User Manual

【Product Name】
Dengue NS1/IgG/IgM Test (GICA)
【Package Specifications】
REF. No. 0320320901 0320320902 0320320903 0320320904
Package Size 10T 25T 50T 100T
【Intended Use】
The Dengue NS1/IgG/IgM Test (GICA) is a lateral flow immunoassay intended for the
simultaneously qualitative detection of Dengue NS1 antigens, Dengue IgG antibodies and IgM
antibodies in serum, plasma or whole blood from patients with fever. It is for professional use only
and is intended to aid to the presumptive diagnosis of primary and secondary Dengue infection.
【Summary and Explanation】
Dengue fever, is an acute mosquito-borne infectious disease, caused by dengue virus with type 1, 2,
3, 4. In addition to causing dengue fever, dengue virus infection can cause Dengue Haemorrhagic
Fever (DHF) or Dengue Shock Syndrome (DSS) in severe cases. Dengue is considered to be the
most serious arthropod-borne viral disease due to the human morbidity and mortality it caused.
The total length of the dengue virus genome is about 11kb, which encodes three structural
proteins(Nucleoprotein C, membrane binding protein M, and envelope protein E) and seven
non-structural proteins(NS1, NS2a, NS2b, NS3, NS4a, NS4b, and NS5). NS1 protein is a highly
conserved non-structural glycoprotein of dengue virus, exists in membrane-bound form and secretory
form, both of which have strong immunogenicity.
NS1 is present at high concentrations in the sera of dengue-infected patients during the early clinical
phase of the disease. And is found from the first day and up to nine days after onset of illness in cases
of primary or secondary dengue infected patients.
Primary Dengue infection is associated with mild to high fever, headache, muscle pain and skin rash.
Immune response includes IgM antibodies produced by 3rd~5th day of symptoms and persist for
30~60 days. IgG antibodies appear the 14th day and persist for life. Secondary infections often result
in high fever and in many cases with haemorrhagic events and circulatory failure. Secondary
infections show that IgG antibodies rise within 1~2 days after the onset of symptoms and induce IgM
response after 20 days of infection.
【Principle】
 For Dengue NS1 antigen test
This test uses a sandwich immunochromatography method for the detection of Dengue NS1 antigens.
The antigens in the sample bind to antibodies conjugated colloidal gold to form antigen-antibody
complexes. The antigen-antibody complexes migrate through nitrocellulose membrane, which can be
captured by immobilized antibodies on test line (T-Line). The red bind on T-Line indicating positive
Dengue NS1 antigens. Under normal test conditions, the control line (C-Line) should be colored to
indicate that the test is effective.
 For IgG and IgM antibody test
This test uses a sandwich immunochromatography method for the detection of Dengue IgG and IgM
antibodies. The IgG and IgM antibodies in the sample bind to antigens conjugated colloidal gold to
form antigen-antibody complexes. The antigen-antibody complexes migrate through nitrocellulose
membrane, which can be captured by the anti-human IgG antibodies or anti-human IgM antibodies
in the test region (G-Line, M-Line). The red bind in the test region (G-Line, M-Line) indicating a
positive result. Under normal test conditions, the control line (C-Line) should be colored to indicate
that the test is effective.

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【Materials Provided】
Specifications Test Cartridge Pipette Sample Diluent User Manual
10T 10 pcs 10 pcs 1 pc 1 pc
25T 25 pcs 25 pcs 1 pc 1 pc
50T 50 pcs 50 pcs 2 pc 1 pc
100T 100 pcs 100 pcs 4 pc 1 pc
【Materials Required But Not Provided】
1. Timer 2. Specimen collection containers 3. Alcohol pads 4. Centrifuge
5. Personal protective equipment (such as face mask, eye goggles, gloves)
【Storage and Shelf Life】
 The test cartridge is stable for 18 months (while sealed in an aluminum foil pouch) if stored at 4
- 30 ℃.
 The sample diluent is stable for 24 months if stored at 4 - 30 ℃.
 After the test cartridge pouch is opened, the test cartridge is stable for only 1 hour. It should be
used immediately after open the package.
 Use the assay kit before the expiration date marked on the box.
【Specimen Collection and Preparation】
 For Whole Blood
Whole blood samples should be collected with specimen collection containers containing
suitable anticoagulant by standard operation of venous puncture.
 For Serum and Plasma
1. Blood samples should be collected by standard operation of venous puncture; after complete
coagulation, the tangible component should be removed by centrifugation.
2. Separate the serum or plasma samples from blood as soon as possible to avoid hemolysis.
3. Lipid layer floating on the upper of the sample should be removed.
【Sample Requirements】
 It is recommended that specimens should be tested immediately. Do not leave samples at room
temperature for more than 3 hours. Otherwise, samples should be stored at 2~8 ℃ for no more
than 72 hours or at -20 ℃ for no more than 30 days after collection.
 Samples with severe hemolysis, hyperlipidemia or jaundiceare not in suggestion.
 Freezing and thawing cycle is permitted only once.
 Samples and test kits must be equilibrated to room temperature (15-30 ℃) prior to testing.
【Operation Step】
Please read the instruction for use carefully before performing the test.
① Remove out the test cartridges from the box, tear the foil bags and place the test cartridges on a
horizontal table.
② Use a pipette to draw the whole blood, serum or plasma sample.
③ Hold the pipette vertically, slowly add one drop (approximately 10 μL) of the whole blood
sample or two drops (approximately 20 μL) of the serum or plasma sample into the two sample wells
respectively.
④ Add 2 drops (approximately 80 μL) of sample diluent into the two sample wells respectively and
start timing. Do not add bubbles and do not handle or move the test cartridge until the test is
complete and ready for reading.
⑤ Wait for 15-20 minutes at room temperature (15-30 ℃) and read the results.
⑥ Dispose of the used test cartridges, pipettes and sample diluent tubes according to your local
regulations and biohazard waste disposal protocol.

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【Quality Control】
A procedural control is included in the test. A colored line appearing in the control region (C-Line) is
considered an internal procedural control. It confirms sufficient specimen volume, adequate
membrance wicking and correct procedural technique.
We also recommend using external controls to confirm the test procedure and to verify proper
performance. Positive and negative controls should be adopted the same assay procedures, and
should give the expected results.
【Interpretation of Test Result】
 Positive :Colored bands appear at both test region (T-Line or G-Line or M-Line) and two control
lines (C-Line).
 Negative: Colored band appear at two control lines (C-Line) only.
 Invalid: No visible colored band appear at any one control line (C-Line), no matter the test
region (T-Line, G-Line and M-Line) is colored or not.

Note:
 This is a qualitative test only and cannot determine the concentration of analyses in the specimen,
so the intensity of color in the test region (T-Line, G-Line and M-Line) may different depending on
the concentration of analyses present in the specimen. Therefore, any shade of color in the test region
should be considered positive.
 It is recommended that the specimen be re-tested using a new kit if the result is invalid.
【Limitations of Test Method】
 This kit is used for the detection of Dengue NS1 antigens, Dengue IgG and IgM antibodies, not
for any other substance.
 The specimen types of this kit are serum, plasma and whole blood samples.
 An incorrect test result may occur if the specimen was collected, extracted or transported
improperly.

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 Reading the test results earlier than 15 minutes or later than 20 minutes may give incorrect
results.
 The test result may be negative if the level of analyses is below the minimum detection level.
 Negative test results do not rule out the possibility of Dengue infection. If the clinical symptoms
persist, a new sample should be draw from the patient in 3~5 days and then should be retested.
 Positive test results do not rule out co-infections with other pathogens.
【Analytical Performance Characteristics】
1. Positive coincidence rate: the enterprise references (P1-1-P1-10, P2-1-P2-10, P3-1-P3-10) coincidence
rate should be 100%.
2. Negative coincidence rate: the enterprise references (N1-N10) coincidence rate should be 100%.
3. Limit of Detection: the enterprise references (L1-1, L1-2, L2-1, L2-2, L3-1, L3-2) should be positive,
L1-3, L2-3 and L3-3 can be positive or negative, the references (L1-4, L1-5, L2-4, L2-5, L3-4, L3-5) should be
negative.
4. Intra-batch repeatability: the enterprise references (J1, J1-2, J2-2, J3-2) should be tested for 20 times
respectively with the same batch of kits, J1 should be negative, J1-2, J2-2 and J3-2 should be positive.
5. Inter-batch repeatability: the enterprise references (J1, J1-2, J2-2, J3-2) should be tested for 10 times
respectively with three batches of kits, J1 should be negative, J1-2, J2-2 and J3-2 should be positive.
6. Analytical specificity
① Cross-reactivity
Evaluation was performed using positive serum samples of different pathogen, this kit is
non-reactive with the following substances.
No. substance Strain No. substance Strain No. substance Strain
1 HKU1 12 Type 1 23 Influenza B virus B/Lee/40
Coxsackie
2 229E 13 Type 2 24
Coxsackie virus A2
Human virus Coxsackie
3 OC43 14 Type 3 25
Coronavirus virus A4
Adenovirus
4 NL63 15 Type 4 26 B1 - conn5
Coxsackie
virus B3 – nancy
5 MERS 16 Type 5 27
(5A1)
6 HCMV HCMV-AD-169 17 Type 7 28 Echovirus 6
7 HSV-1 - F(3A20) 18 Type 55 29 Echovirus Echovirus 9
HSV
8 HSV-2 - MS(4A6) 19 H1N1 30 Enterovirus 71
Polio virus Rhinovirus -
9 Polio virus 20 H3N2 31 Rhinovirus
-sabin(3A4) Influenza A RV21
10 Corona FCV(3A2) 21 H5N1 32 Respiratory Type A
11 virus FIP(2A4) 22 H7N9 33 syncytial virus Type B

② Interference
There was no significant interference effect on this kit from these substances.
No. Interference materials Conc. No. Interference materials Conc.
1 Bilirubin 30 mg/dL 3 Hemoglobin 200 mg/dL
2 Lipid 1500 mg/dL 4 HSA 8 g/dL

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7. Clinical performance evaluation
This kit has demonstrated the following clinical performance results.
Reference reagent
For Dengue NS1 antigens
Positive Negative Total
Dengue Positive 194 4 198
NS1/IgG/IgM Test Negative 6 246 252
(GICA) Total 200 250 450

Reference reagent
For Dengue IgG and IgM antibodies
Positive Negative Total
Dengue Positive 196 5 201
NS1/IgG/IgM Test Negative 4 245 249
(GICA) Total 200 250 450

Dengue NS1/IgG/IgM Test (GICA)

Sensitivity (positive coincidence rate) Specificity (negative coincidence rate)

Dengue NS1 antigen 97.0% (95% CI: 93.61% - 98.62%) 98.4% (95% CI:95.96% - 99.38%)
Dengue IgG and
98.0% (95% CI: 94.97% - 99.22%) 98.0% (95% CI:95.40% - 99.14%)
IgM antibody

【Warnings and Precautions】

 For in vitro diagnostic use only. For professional use only.
 The test cartridges and pipettes in this kit are intended for single use only.
 Do not use the test kit if the pouch is damaged.
 Wear personal protective equipment when handing specimen or used kit components.
 Do not mix reagent of different lots or those for other products.
 The used test cartridges and pipettes may be potential infectious. Proper handing and disposal
methods should be established in accordance with local regulatory requirements.
 The reagent solution contains a salt solution, so flush with copious amounts of water if the
solution contacts the skin or eyes.
 Any serious incident that has occurred in relation to the device shall be reported to the
manufacturer and the competent authority.
【Literature References】
[1]Dussart P. etc. al, 2006. Evaluation of an enzyme immunoassay for detection of dengue virus NS1
antigen in human serum. Clin. Vaccine Immunol. 13:1185-1189.
[2]Chung K. M. etc. Al, 2003. Antibodies against West Nile Virus nonstructural protein NS1 prevent
lethal infection through Fc gamma receptor-dependent and -independent mechanisms. Journal of
Virology, 2006(3).
[3]Gubler, DJ. 1994. Perspectives on the prevention and control of dengue hemorrhagic fever.
Kaohsiung J Med Sci, 10: 15-18.

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【Symbol Explanation】
Interpretation of Interpretation of Interpretation of
Symbol Symbol Symbol
symbols symbols symbols
manufacturer Do not re-use CE marking
Authorized
representative in the Consult instructions
Catalogue number
European for use
Community
In vitro diagnostic
Date of manufacture Caution
medical device
Contains sufficient
Use-by date Temperature limit
for <n> tests
Batch code
【Manufacturer's Information】
Shenzhen Lifotronic Technology Co., Ltd.
Unit A, 4th Floor, Building 15, Yijing Estate, No.1008 Songbai Road, Nanshan
District, Shenzhen City, Guangdong Province, 518055, P.R.China
Mail：service@lifotronic.com;

Umedwings Netherlands B.V.

Treubstraat 1, 2288EG, Rijswijk, The Netherlands
Tel.+31(0) 642758955
Mail:ar@umedwings.eu
【Version and Modification Date】
Version: A0
Modification Date: 2020.03.05

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