DRG JE (Japanese Encephalitis) IgM Capture Elisa Kit

PRINCIPLE OF THE TEST 
The JE (Japanese Encephalitis) IgM Capture ELISA consists of one enzymatically amplified "two-step" sandwich-type immunoassay.
In this assay, JE Negative Control (represents non-reactive serum), JE IgM Positive Control (represents reactive serum), and unknown serum samples are diluted with Sample Dilution Buffer, then incubated in microtitration wells which have been coated with anti-human IgM antibodies. This is followed by incubation with both JEV-derived recombinant antigen (JERA) and Normal Cell Antigen (NCA) separately. After incubation and washing, the wells are treated with a JERA-specific antibody labeled with the enzyme horseradish peroxidase (HRP). After a third incubation and washing step, the wells are incubated with the tetramethylbenzidine (TMB) substrate. 
An acidic stopping solution is then added and the degree of enzymatic turnover of the substrate is determined by absorbance measurement at 450 nanometers. Above a certain threshold, the ratio of the absorbencies of the JERA and the control wells accurately determines whether antibodies to JEV are present.

SPECIMEN COLLECTION AND PREPARATION 
Human serum must be used with this assay. Whole blood or plasma cannot be tested directly. 
Note: CSF can be used. However, our kit has not been tested or optimized with CSF. Before using this kit, one has to optimize the CSF system.
Remove serum from the clot of red cells as soon as possible to avoid hemolysis. 
Testing should be performed as soon as possible after collection. Do not leave sera at room temperature for prolonged periods.
Serum should be used and the usual precautions for venipuncture should be observed. The samples may be stored at 2 °C - 8 °C for up to 7 days. To maintain long-term longevity of the serum, store at -20 °C or lower. Avoid repeated freezing and thawing of samples. 
Frozen samples should be thawed to room temperature and mixed thoroughly by gentle swirling or inversion prior to use. Always quick spin before use. 
If sera are to be shipped, they should be packed in compliance with Federal Regulations covering transportation of infectious agents. 
Do not use sera if any indication of growth is observed.

QUALITY CONTROL 
Each kit contains positive and negative controls to ensure assay performance. 
The negative and positive controls are intended to monitor for substantial reagent failure. The positive control will not ensure precision at the assay cut-off. The test is invalid and must be repeated if the ISR values of either of the controls do not meet the specifications. 
Acceptable Immune Status Ratio (ISR) values for these controls are found on the specification table below. If the test is invalid, patient results cannot be reported. 
Quality control requirements must be performed in conformance with local, state, and/or federal regulations or accreditation requirements and your laboratory’s standard Quality Control procedures. It is recommended that the user refer to CLSI C24 and 42 CFR 493.1256 for guidance on appropriate QC practices. 
The results below are given strictly for guidance purposes only. 
Applicable for raw spectrophotometric readings only. Do not subtract any background.

TEST PROCEDURE
Bring all kit reagents and specimens to room temperature (20 °C - 25 °C) before use. Thoroughly mix the reagents and samples before use by gentle inversion.