This test applies the Capture ELISA principle and uses mouse anti-human IgM (Anti-μ chain) monoclonal antibody-coated micro-well plate as the solid carrier. It can capture herpes simples virus (HSV)-I IgM antibodies in a sample, and uses horseradish peroxidase-labelled HSV-I antigens as the tracer and TMB coloration system. If a sample contains HSV-I IgM antibodies, they will react with labelled antigens, otherwise, no color will show. Use a micro-plate reader to test the sample absorbance (OD value) in order to determine the presence of HSV-I IgM antibodies in the human serum or plasma. If the sample’s tested OD value is lower than the cutoff value, then the test is determined as negative, whereas if the OD value is higher than the cutoff value, the the test is determined as positive. Therefore this kit can detect serum samples that contain herpes simples virus type I IgM antibodies.
1 Temperature Balancing: Take the kit out from the refrigerate and wait until it reaches the room temperature.
2 Adding Diluent: Dilute the concentrated washing solution 20 times with purified or deionized water and shake well.
3 Adding Sample: Leave an empty well without adding any liquid (including any enzyme). Also leave 3 wells for negative control and 2 wells for positive control. Add 20μl each of above mentioned controls to their corresponding wells and add 20μl of testing sample to the rest of the wells.
4 Adding Enzyme: Add 100ml enzyme conjugate to each well (apart from the control wells) and mix well by oscillating.
5 Incubation: Cover the microplate with cover membrane and incubate for 60 minutes in over or water bath set for 37℃±1℃.
6 Washing: Discard all the solution from the microplate fill up each will with washing solution. Wait for 30 seconds and discard all the solution from the microplate. Repeat the washing step 5 times and pat dry the microplate with a paper towel. Alternatively, use a micro plate washer to wash the plate 5 times and pat dry the plate with a paper towel.
7 Coloration: Add 50ml each of substrate A and B to each well (including the empty control wells), cover the microplate with a cover membrane and mix well by oscillating. Incubate at 37℃±1℃ in the dark for 15 minutes.
8 Stop: Add 50μl of stop solution to each well mix well by oscillating.
9 Result determination: Use a microplate reader（450/630nm or 450nm）to read the result from each well (the result must be determined within 30 minutes after the adding of stop solution).
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