Enterovirus 71 (EV71)-IgM Ab Cassette

INTENDED USE

This test kit is used to qualitatively detect the Enterovirus 71 (EV71)-IgM antibody in the human blood, serum or plasma samples. It is a diagnostic reagent of EV in early infection and is used in clinical diagnosis. It is one of the Picornaviridae virus (Enterovirus, EV) genus, group A and has obvious properties of dermatotropic and neurotropic. Being infected by EV71 can cause hand-foot-mouth disease, Aseptic Meningitis, Herpes Pharyngitis, and encephalitis, and the paralysis of poliomyelitis and recessive infection in other cases. A human being is known as a natural host of EV71. It is mainly infected by the fecal-oral transmission. Its epidemic season is summer and autumn, but it can infect all year. Human is the generally susceptible of EV71, while the infants and young children are at more risk.

PRINCIPLES OF THE TESTS

This test kit applies the technology of colloidal gold immunochromatography for the qualitative test of EV71- IgM antibody present in the serum, whole blood or plasma samples. The EV71-IgM antibodies in a positive sample can combine with the colloidal gold labeled mouse-anti-human IgM immobilized on a conjugate pad to form immune complex, and move along the membrane strip by chromatography. Some of the complex is caught by the recombinant EV71-Ag pre-coated on the T line to form "gold labeled mouse-anti-human IgM- EV71 IgM Ab- EV71 Ag" and develop a color. The remaining free complex is caught by the goat-anti-mouse IgG antibody coated on the C line to develop a color. As for negative samples, in which there are no EV71- IgM antibodies, cannot form immune complexes and develop color on the T line, and colorate on the C line only.

MAIN CONSTITUENT

• Test card (Coating the recombinant EV71 VP1-VP3 antigen on T-line, the goat-anti-mouse IgG antibody on C-line): 40 test cards/ box

• The sample diluents: 1 bottle (5 ml)

• Product instruction: 1

STORAGE CONDITIONS AND THE PERIOD OF VALIDITY

• Can be stored for 24 months in a dry place at 4° to 30℃

• Use test card within 1 hour once opened to atmosphere where the humidity is below 60%, or use it immediately if the humidity is higher.

SAMPLE REQUEST

• Serum and plasma samples are collected from the venous blood by the conventional method. Plasma sample: add 100ul heparin solution (1%) to 5 to 10ml blood; or sodium citrate solution (3.8%) to plasma according to the proportion of 1:9; or EDTA solution (15%) 0.04ml to 5ml plasma.

• Serum and plasma samples can be stored at 4℃ if test is done within 5 days or else store at -20 ℃ and not more than 3 times of freeze-thaw.

• The whole blood sample should be stored at 2 to 8℃ and tested within 3 days, and should not be frozen.

• The test result is invalid for hemolytic sample.

TEST PROCEDURE

• Test preparation: 10μL, 50μL, 100μL micropipettes, and matched tips

• Test process: Make sure that before the test the temperature of the kit and the sample should be the same with the room temperature. Place the test card on a horizontal dry work surface. Add 10μL serum or plasma sample into the sample well or 50μL whole blood then add 100μL of sample dilution immediately. Observe the result in 15 to 20 minutes after the serum or plasma samples are added. The observation is invalid after 20 minutes.

TEST RESULTS EXPLAIN

The color developed on C line only: negative; color developed on both C line and T line: positive; color developed on T line only: invalid; color not developed on both C line and T line: invalid.

Note: Re-detection if the detected result is invalid. The invalid test cards should be dealt with as infectious pollutants and the temperature of the kit and sample should be the same with room temperature before pre-detection.

LIMITATION

1. It was vulnerable to the visual error or the subjective judgment factors duplicate the detection when the stripe color is not obvious.

2. The detection card is one of the assistant diagnostic methods; its test results are only for the reference and should not be the sole base for clinical diagnosis and its treatment. The positive results should be further verified by other methods also. Due to the limit of its detection sensitivity, the negative results should also be observed because the concentration of the antibodies is lower than the analysis sensitivity. It should combine with the clinical diagnosis, medical history, and other detection methods.

3. During the early stage of infection, IgM may not be generated or may be in low titer. It may give negative results. Suggest the patients review the test in 7 to 14 days and do the parallel detection to the old samples at the same time, so as to confirm whether the serological positive or titer increased significantly.

4. The results will be unreliable for patients with impaired immune function or immunosuppressive treatment.

5. Positive results will not only occur in the primary infection but also in the secondary infection.

6. This test kit is a qualitative test and cannot be used for determining the antibody levels.

7. It is used for the test of serum or plasma samples only, not for saliva, urine or any other body fluids testing.

PRODUCT PERFORMANCE INDICATORS

Positive coincidence rate: to internal reference (+/+) =12/12; Negative coincidence rate (-/-) =12/12; precision (n=10): positive for all tests, and develop color equably; Minimum detection limit: positive end point is not less than 1:8 or 1:16 dilution of positive reference L1. The result must be negative to reference L2. The samples which is positive with toxoplasma IgM antibody (S/C value: ≤ 13.65), rubella virus IgM antibody (S/C value: ≤ 11.42), cytomegalovirus IgM antibody (S/C value: ≤ 13.41), herpes simplex virus II IgM antibody (S/C value: ≤ 12.53), Toxoplasma IgG antibody (S/C value: ≤ 13.45), rubella virus IgG antibody (S/C value: ≤ 10.39), cytomegalovirus IgG antibody (S/C value: ≤ 13.15), herpes simplex virus II IgG antibody (S/C value: ≤ 14.28), hepatitis B virus surface antigen (S/C value: ≤ 13.57), hepatitis C virus surface antigen positive (S/C value: ≤ 14.75), CVA16 antibody (S/C value: ≤ 14.28), HIV (S/C values: ≤ 11.37), HEV-IgM (S/C values: ≤ 12.15), rheumatoid factor (≤ 54IU/ml), antinuclear antibody (≤ 1:640), Vibrio cholera, Salmonella etc. cannot affect results. There is impact in test when the lipid content of the sample is higher than 6mmol / L and the Bilirubin level is higher than 40μmol / L.

PRECAUTIONS

• When the kit test result is positive use other methods also for confirmation

• Avoid the test card being exposed in the air for a long time since it can absorb moisture that can affect the test results. Use the test card within 1 hour once opened to the atmosphere and humidity is below 60%, or use it immediately if the humidity is higher.

• The degree of coloration on the test line does not connect to the antibody titers in the test sample.

• The color of C line is likely to abate when the content of the virus antibodies is extremely high in the sample. It is a normal phenomenon.

• Be attentive to the potential biological risks. Wear necessary protective equipment, and deal with the waste as infectious material.