ELISA Kit for Antibody IgG to Hepatitis E Virus (HEV)
Contents
1. 4 BIOCHAIN www biochain com ELISA Kit for Antibody IgG to Hepatitis E Virus HEV Catalog No Z7010008 NAME AND INTENDED USE ELISA Kit for Antibody IgG to Hepatitis E Virus HEV is an in vitro enzyme immunoassay for the detection of Anti HEV antibodies IgG in human serum or plasma It is for diagnosis of early infection and epidemic survey PRINCIPLE This kit uses indirect ELISA method to detect anti HEV antibodies IgG in serum or plasma The purified activated recombinant HEV antigen ORF2 ORF3 is coated on the multi wells plate The HRP conjugated mouse anti human IgG r chain monoclonal antibody serves as tracer TMB is substrate for HRP The enzyme reaction with substrate TMB produces a color change and the intensity of the absorbance at 450 nm indicates the presence or absence of Anti HEV antibodies IgG in the sample The test is specific sensitive reproducible and easy to operate It is for blood screen of HEV infection INSTRUMENT 8 x 12 wells plate reader and washer MATERIALS PROVIDED 96 tests 1 Antigen Coated Microwell Plate 1 block 96wells 2 Sample Diluent 1 bottle 12ml 3 Negative Control Serum 1 vial 1ml 4 Positive Control Serum 1 vial 1ml 5 Concentrated Wash Buffer 20x 1 bottle 50ml 6 Enzyme Conjugant 1 bottle 12ml 7 Substrate A 1 bottle 6ml 8 Substrate B 1 bottle 6ml 9 Stop Solution 1 bottle 6ml 10 Plastic Bag 1 bag 11 Seal Paper 2 pieces 12 Manual 1 eac2. mix thoroughly 8 Cover wells with seal paper incubate for 20 minutes at 37 C 9 Repeat step 6 10 Add 50 ul substrate A and B respectively to each well mix gently protected from light and lay aside for 10 minutes at 37 C 11 Add 50 ul of stop solution into each well to stop the reaction including blank well 12 Measure the absorbance at 450 nm against the blank or measure the absorbance at 450 nm 630 690 nm INTERPRETATION OF RESULTS Colorimetric Method Cut Off Value calculation COV 0 1 the average OD of negative controls if the average OD of negative controls is below or equal to 0 05 calculate it as 0 05 Positive ODas0 of sample COV Negative OD 50 of sample lt COV LIMITATION The testing is for qualitative and assistant diagnosis Confirmation of infection should refer to the clinical and other diagnosis QUALITY CONTROL If the OD of positive controls is not below 1 0 OD of negative is not higher than 0 1 the assay result is validated Otherwise repeat the test PRECAUTIONS 1 The samples should be fresh avoid hemolysis bacteria growing and repetitive freeze and thaw 2 Do not interchange reagents between kit lots The seal paper can t be used repeatedly 3 Mix reagents well before use If crystal form in certain reagents such as wash buffer it can be used without problems after warm up and mix well 4 Follow instruction exactly during assay especially in temperature and t
3. h SAMPLE COLLECTION AND PRESERVATION Blood serum samples are routinely prepared form vein Blood plasma sample are routinely prepared with routine amount of anticoagulant such as heparin or sodium citrate Sample can be stored at 4 C if tested within five days Sample can be stored at 20 C at least for 3 months Avoid hemolysis and repetitive freeze and thaw of samples Samples with cloud or precipitation should be centrifugated or filtered before test Prevent serum from bacteria contamination during collection and storage TEST PROCEDURE 1 Bring ELISA Kit for Antibody IgG to Hepatitis E Virus HEV all reagents and samples to room temperature before use approximately 30 minutes 2 Dilute concentrated wash buffer 1 20 with ddH20 3 For each test set one blank two positive and three negative controls Add 100 ul positive and negative control serum into positive and negative control wells respectively without sample diluent 4 Add 100 ul sample diluent in each test wells then at 10 ul test serum into test wells Pipet up and down to mix the samples well 5 Cover wells with seal paper incubate for 20 minutes at 37 C 6 Discard the liquid in all wells and fill the wells with wash solution 350ul per well Lay aside for 15 seconds discard the liquid in all wells and fill the wells with wash solution Repeat 5 times and dry wells after last wash 7 Add enzyme conjugant 100 ul into the wells except the blank well
4. ime for reactions All pipetting devices should be used with care and calibrated regularly following the manufacturer s instructions 5 Put the remained reagents to the sealed pouch and return to 2 8 C in time 6 To prevent cross contamination wear gloves and working suits throughout the procedure and execute the disinfection and isolation regulations strictly Dispose of all samples and materials used to perform the test The 5 0g L liquid sodium hypochlorite solution or 121 C high pressure steam may be used to disinfect samples and materials before disposal The positive control serum in the kit has been inactivated already STORAGE AND STABILITY Store the kit at 2 8 C EXPIRATION The shelf life is 12 months from the receiving date This Kit is for Research Use Only
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