Document No. : INS-DG-EN (Rev.

00)
Revision date : July 5, 2017

WARNINGS AND PRECAUTIONS

For in vitro diagnostic use only.
Carefully follow the instructions and procedures described in this
‘Instruction for use’.

Dengue IgG/IgM
Use only fresh samples and avoid direct sunlight.
Lot numbers of all the test components (cartridge, ID chip and
detection buffer, diluent) must match each other.
Do not interchange the test components between different lots
or use the test components after the expiration date, either of
INTENDED USE which might yield misleading of test result(s).
ichroma™ Dengue IgG/IgM is a fluorescence Immunoassay (FIA) for Do not reuse. A detection buffer tube should be used for
the qualitative determination of IgG/IgM antibodies against dengue processing one sample only. So should a cartridge.
virus in human whole blood/serum/plasma. It is useful as an aid in The cartridge should remain sealed in its original pouch before
screening of Dengue virus infection. use. Do not use the cartridge, if is damaged or already opened.
For in vitro diagnostic use only. Frozen sample should be thawed only once. For shipping,
samples must be packed in accordance with the regulations.
Sample with severe hemolytic and hyperlipidemia cannot be
INTRODUCTION used and should be recollected.
Dengue virus (DENV), a mosquito-borne virus, phylogenetically Just before use, allow the cartridge, detection buffer and sample
belongs to the genus Flavivirus, including Zika virus, West Nile virus, to be at room temperature for approximately 30 minutes.
yellow fever virus, and Japanese encephalitis virus. Dengue virus ichroma™ Dengue IgG/IgM as well as the instrument for
comprises of 4 serotypes distinct in the infection tendency and immune ichroma™ tests should be used away from vibration and/or
responses. Dengue virus infection causes clinically a wide range of magnetic field. During normal usage, it can be noted that
human diseases from mild Dengue Fever (DF) to severe Dengue instrument for ichroma™ tests may produce minor vibration.
Hemorrhagic Fever (DHF) or Dengue Shock Syndrome (DSS).1) Several Used detection buffer tubes, pipette tips and cartridges should
lines of evidences show that secondary Dengue virus infection, with be handled carefully and discarded by an appropriate method in
different serotypes from the primary infection, is relevant to severe accordance with relevant local regulations.
Dengue diseases.2) The immune response to primary or secondary An exposure to larger quantities of sodium azide may cause
virus infection varies. In the case of primary infection, specific IgM is certain health issues like convulsions, low blood pressure and
higher titre during 4-10 days after onset of illness than IgG. IgG heart rate, loss of consciousness, lung injury and respiratory
response become permanent for whole life of patient with primary failure.
infection. During secondary infection, in contrast, the titre of virus- ichroma™ Dengue IgG/IgM will provide accurate and reliable
specific IgG is higher than IgM titre in whole of serological period.2,3) results subject to the following conditions.
Although there are many types of serological diagnostic reagents - Use ichroma™ Dengue IgG/IgM should be used only in
including enzyme-linked immunosorbent assay (ELISA) or conjunction with instrument for ichroma™ tests.
immunofluorescent assays (IFA), development of simultaneous and - Any anticoagulants other than EDTA, heparin, sodium citrate
accurate detection kit of IgG and IgM is required.4) should be avoided.

PRINCIPLE STORAGE AND STABILITY

The test uses a sandwich immunodetection method; granulized The cartridge is stable for 20 months (while sealed in an
detector antigen in cartridge binds to antibody in the sample, forming aluminum foil pouch) if stored at 4-30 °C.
antibody-antigen-antibody complexes, and migrates onto The detection buffer is stable for 20 months if stored at 2-8 °C.
nitrocellulose matrix to be captured by the other immobilized-antigen The diluent dispensed in a vial is stable for 20 months if stored at
on the test strip. 2-8 °C.
The more antibody in sample forms the more antigen-antibody After the cartridge pouch is opened, the test should be
complex, which leads to stronger intensity of fluorescence signal on performed immediately.
detector antigen, which is processed by the instrument for ichroma™
tests to show ‘dengue IgG/IgM positive’ in sample. LIMITATION OF THE TEST SYSTEM
The test may yield false positive result(s) due to the cross-
COMPONENTS reactions and/or non-specific adhesion of certain sample
ichroma™ Dengue IgG/IgM consists of ‘Cartridges’, ‘Detection Buffer components to the capture/detector antigens.
Tubes’, ‘Diluent vial’ and an ‘ID chip’. The test may yield false negative result. The non-responsiveness
The cartridge contains a test strip, the membrane which has anti- of the antibodies to the antigens is most common where the
human IgM and anti-human IgG at each test lines respectively, epitope is masked by some unknown components, so as not to
while Nus antigen at the control line. be detected or captured by the antibodies. The instability or
Each cartridge is individually sealed in an aluminum foil pouch degradation of the antigen with time and/or temperature may
containing a desiccant. 25 sealed cartridges are packed in a box cause the false negative as it makes antibody unrecognizable by
which also contains an ID chip. the antigens.
The detection buffer contains granules. There are contain anti- Other factors may interfere with the test and cause erroneous
Dengue antigen fluorescence conjugate, anti-Nus fluorescence results, such as technical/procedural errors, degradation of the
conjugate, bovine serum albumin (BSA) as a stabilizer and sodium test components/reagents or presence of interfering substances
azide as a preservative in phosphate buffered saline (PBS). in the test samples.
25 detection buffer tubes are packed in an aluminum foil pouch. Any clinical diagnosis based on the test result must be supported
The diluent contains a detergent and bovine serum albumin (BSA) by a comprehensive judgment of the concerned physician
as a stabilizer and sodium azide in sodium borate buffer as a including clinical symptoms and other relevant test results.
preservative. The diluent is dispensed in a vial.

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Document No. : INS-DG-EN (Rev. 00)
Revision date : July 5, 2017

MATERIALS SUPPLIED (The sample mixture must be used immediately.)

4) Pipette out 75 µL of a sample mixture and dispense it into the
REF CFPC-60 DB well on the cartridge.
5) Leave the sample-loaded cartridge at room temperature for 12
Components of ichroma™ Dengue IgG/IgM minutes.
Cartridge Box: Scan the sample-loaded cartridge immediately when the
- Cartridges 25 incubation time is over. If not, it will cause inexact test result.
6) To scan the sample-loaded cartridge, insert it into the cartridge
- ID Chip 1
holder of the instrument for ichroma™ tests. Ensure proper
- Instruction For Use 1 orientation of the cartridge before pushing it all the way inside
Aluminum Pouch containing Detection Buffer Tubes the cartridge holder. An arrow has been marked on the
- Detection Buffer tubes 25 cartridge especially for this purpose.
Dilution Buffer Vial Pouch 7) Tab the ‘Start’ icon on the screen.
- Diluent Vial 1 8) Instrument for ichroma™ tests will start scanning the sample-
loaded cartridge immediately.
MATERIALS REQUIRED BUT SUPPLIED ON DEMAND 9) Read the test result on the display screen of the instrument for
ichroma™ tests.
Following items can be purchased separately from ichroma™ Dengue
IgG/IgM.
INTERPRETATION OF TEST RESULT
Please contact our sales division for more information.
Instrument for ichroma™ tests Instrument for ichroma™ tests calculates the test result
- ichroma™ II REF FPRR021 automatically and displays “IG value: Positive / Negative /
Indeterminate, IM value: Positive / Negative / Indeterminate”.
ichroma™ Printer REF FPRR007
Ancillary value is served in the form of a cut-off index (COI).
Cut-off index
SAMPLE COLLECTION AND PROCESSING Result Note
(COI)
Negative for Dengue
The sample type for ichroma™ Dengue IgG/IgM is human whole ≤ 0.9 No need to additional test.
IgG Ab / IgM Ab
blood/serum/plasma. Need to retest. If test results are
It is recommended to test the sample within 24 hours after shown ‘Negative’ or ‘Indeterminate’
collection. > 0.9,
Indeterminate repeatedly, these samples are
< 1.1 considered dengue IgG/IgM antibody
The serum or plasma should be separated from the clot by
centrifugation within 3 hours after the collection of whole blood. negative.
Samples may be stored for up to a week at 2-8 °C prior to being Positive for Dengue
≥ 1.1 Need to confirmation test.
IgG Ab / IgM Ab
tested. If testing will be delayed more than a week, samples
should be frozen at -20 °C.
QUALITY CONTROL
Samples stored frozen at -20 °C for 2 months showed no
performance difference. Quality control tests are a part of the good testing practice to
However, the whole blood sample should not be kept in a freezer confirm the expected results and validity of the assay and should
in any case. be performed at regular intervals.
Once the sample was frozen, it should be thawed one time and The control tests should be performed immediately after opening
only for test, because repeated freezing and thawing can result a new test lot to ensure the test performance is not altered.
in the changed test values. Quality control tests should also be performed whenever there is
Samples containing precipitates must be clarified by any question concerning the validity of the test results.
centrifugation. Control materials are not provided with ichroma™ Dengue
IgG/IgM. For more information regarding obtaining the control
materials, contact Boditech Med Inc.’s Sales Division for
TEST SETUP assistance.
Check the contents of ichroma™ Dengue IgG/IgM: Sealed (Please refer to the instruction for use of control material.)
Cartridge, Detection Buffer Tubes, Diluent Vial and ID Chip.
Ensure that the lot number of the cartridge matches that of the PERFORMANCE CHARACTERISTICS
ID chip, the detection buffer as well as the diluent. Analytical Sensitivity
Keep the sealed cartridge (if stored in refrigerator), the detection - Limit of Detection (LOD)
buffer tube and the diluent at room temperature for at least 30 To determine the limit of detection (LOD), the panel was tested
minutes just prior to the test. Place the cartridge on a clean, dust- by serial dilution with ichroma™ Dengue IgG/IgM
free and flat surface.
Dengue IgG positive panel
Turn on the Instrument for ichroma™ tests.
Insert the ID Chip into the ID chip port of the Instrument for Commercial ELISA Ichroma™ Dengue IgG/IgM
ichroma™ tests. Dilution ISR of
Press the ‘Select’ button on the Instrument for ichroma™ tests. Results COI CV (%)
Factor ELISA
(Please refer to the ‘Instrument for ichroma™ tests Operation 20 8.24 Positive 18.45 3.4
Manual’ for complete information and operating instructions.) 2-1 6.56 Positive 10.21 5.1

TEST PROCEDURE 2-2 4.84 Positive 6.98 8.3

2-3 3.84 Positive 5.43 4.3
1) Transfer 10 µL of sample (whole blood/serum/plasma/
2-4 2.84 Equivocal 3.85 13.6
control) using a pipette to the sample well on the cartridge.
2) Transfer 150 µL of the diluent using a pipette to the detection 2-5 2.18 Equivocal 2.16 10
buffer tube. 2-6 2.14 Equivocal 1.01 5.8
3) Dissolve the granulized detection buffer thoroughly by tapping 2-7 2.01 Equivocal 0.98 44.2
10 times and pipetting 10-20 times. 2-8 1.07 Negative 0.99 19.7

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Document No. : INS-DG-EN (Rev. 00)
Revision date : July 5, 2017

Between lot Between person Between day Between site

Dengue IgM positive panel Dengue
Positive/ Positive/ Positive/ Positive/
IgG/IgM
FDA-cleared MAC ELISA ichroma™ Dengue IgG/IgM Number Positive Number Positive Number Positive Number Positive
Cal
Dilution ISR of of test of test of test of test
Results COI CV (%)
Factor ELISA Negative 0/10 0% 0/5 0% 0/5 0% 0/5 0%

2 0
10.66 Positive 7.84 1.1 High 10/10 100 % 5/5 100 % 5/5 100 % 5/5 100 %
Mid 10/10 100 % 5/5 100 % 5/5 100 % 5/5 100 %
2-1 4.6 Positive 1.97 7.1
Low 10/10 100 % 5/5 100 % 5/5 100 % 5/5 100 %
2-2 3.02 Positive 0.98 7.4
2-3 1.75 Equivocal 1.01 25.1 Comparability with reference product
2-4 0.38 Negative 0.87 35.2 Commercial IgG ELISA result
-5
2 0.13 Negative 0.89 28.2 Positive Equivocal Negative Total

* In the case of Dengue IgG panel, ichroma™ Dengue IgG/IgM Positive 40 0 0 40

shows more sensitive analytical reactivity by 16 times than result ichroma™ Equivocal 0 0 0 0
of reference ELISA. Dengue IgG/IgM Negative 0 0 50 50
* In the case of Dengue IgM panel, ichroma™ Dengue IgG/IgM Total 40 0 50 90
shows similar sensitivity to result of reference ELISA. FDA-cleared MAC-ELISA result
Positive Equivocal Negative Total
Analytical Specificity
Positive 26 0 1 27
- Cross-reactivity
ichroma™ Equivocal 0 0 0 0
There was no false positive result from 169 samples containing
potentially interfering substances with the ichroma™ Dengue Dengue IgG/IgM Negative 1 0 49 50
IgG/IgM test. The overall specificity was 100 %. Total 27 0 50 77
Clinical ichroma™ Dengue IgG/IgM
- IgG sensitivity = 40/40 x 100 = 100 %
category Number of samples Negative Positive
- IgG specificity = 50/50 x 100 = 100 %
ZIKV 17 17 0
- IgM sensitivity = 25/26 x 100 = 96.15 %
CMV 20 20 0
- IgM specificity = 48/49 x 100 = 97.95 %
EBV 20 20 0
HAV 28 28 0
HCV 15 15 0
REFERENCES
HBV 21 21 0 1. Evaluation of diagnostic test: Dengue. Rosanna W. P. et al., Nature,
ANA 23 23 0 2010
RF 25 25 0
2. Immunoglobumin G (IgG) to IgM ratio in secondary adult dengue
infection using samples from early days of symptoms onset.
Total 238 238 0
Cucunawangsih et al., BMC Infectious Diseases, 2015
- Interference 3. Dengue Viraemia Titer, Antibody Response Pattern and Virus
There was no significant interference from these material with Serotype Correlate with Disease Severity: Vaughn. D. W. et. al.,
the ichroma™ Dengue IgG/IgM test.
Journal of Infectious Diseases. 2000
Materials Concentration
4. Current Global Status of Dengue Diagnostics: Miranda D. S. et. al.,
Heparin 100,000 U/L
Journal of Advance in Biology and Biotechnology, 2015
EDTA 4 μM
Sodium citrate 0.085 M Note: Please refer to the table below to identify various symbols.
Bilirubin 200 μM
Hemoglobin 2 g/L
Triglycerides 1.5 mg/mL
Cholesterol 20 mM
Albumin 60 mg/mL

Precision
- Between Lot
One person tested three different lots of ichroma™ Dengue
IgG/IgM, ten times at each concentration of the control standard.
- Between person
Three different persons tested same lot of ichroma™ Dengue
IgG/IgM, five times at each concentration of the control standard.
- Between day
One person tested same lot of ichroma™ Dengue IgG/IgM during
three days, five times at each concentration of the control
standard.
- Between site
One person tested same lot of ichroma™ Dengue IgG/IgM at
three different sites, five times at each concentration of the
control standard.

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Document No. : INS-DG-EN (Rev. 00)
Revision date : July 5, 2017

For technical assistance; please contact:

Boditech Med Inc.’s Technical Services
Tel: +82 33 243-1400
E-mail: sales@boditech.co.kr

Boditech Med Incorporated

43, Geodudanji 1-gil, Dongnae-myeon,
Chuncheon-si, Gang-won-do, 24398
Republic of Korea
Tel: +(82) -33-243-1400
Fax: +(82) -33-243-9373
www.boditech.co.kr

Obelis s.a
Bd. Général Wahis 53,
1030 Brussels, BELGIUM
Tel: +(32) -2-732-59-54
Fax: +(32) -2-732-60-03
E-Mail: mail@obelis.net

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