The Hepatitis B Virus E Antigen (HBeAg) ELISA Kit is a qualitative in vitro enzyme immunoassay for the detection of HBeAg in human serum or plasma.
Principle of the Assay:Purified Anti-HBe is coated on the solid phase of multi-wells. Serum samples and Anti-HBe-HRP conjugate are added to coated wells. After incubation, if HBeAg is present in the sample, a complex of Anti-HBe-HBeAg-Anti-HBe-HRP will form. Wells are washed to remove other unbound serum components and incubated with substrate (TMB) to form a colored product. The absorbance at 450nm is measured to indicate the presence or absence of HBeAg in the sample. The test is sensitive, reproducible and easy to operate.
Kit Components:H1910-40K1: Microtiter Plate, 1x96 wells. Coated with Anti-HBeH1910-40K2: Enzyme Conjugate, 1x6.5mlH1910-40K3: Positive Control Serum, 1x1mlH1910-40K4: Negative Control Serum, 1x1mlH1910-40K5: Wash Buffer, 20X, 1x50ml. Dilute 1:20 prior to use
H1910-40K6: Substrate A, 1x8.5mlH1910-40K7: Substrate B, 1x8.5mlH1910-40K8: Stop Solution, 1x8.5ml
Storage And Stability:Store all kit components at 4°C. Kit is stable for 6 months after receipt.
Test Procedure:1. Set one blank, two positive controls and two negative controls for each assay. Add 50ul serum sample, positive control serum and negative control serum into the coated wells.
2. Add one drop (approximately 50ul) of Enzyme Conjugate into the wells, except the blank wells. Mix thoroughly, cover the wells with seal paper, and incubate for 30-60 minutes at 37°C.
3. Wash the wells. Manual Wash Procedure: Remove the liquid completely from the coated wells. Fill the wells with wash solution and then remove the liquid completely. Repeat 5 times
Automatic Wash Procedure: Select the automatic operation of washing to 5 times; ensure liquid has been removed completely after the last wash.
4. Add one drop (approximately 50ul) of Substrate A and B respectively to each well, seal the plate with seal paper, and incubate for 10-15 minutes at 37°C. Avoid exposure to light.
5. Add one drop (approximately 50ul) of Stop solution into each well and then mix thoroughly to terminate the reaction. Measure the absorbance at 450nm against the blank or measure the absorbance at 450nm/630-690nm.
Interpretation of Results:Cut-off Value (COV) Calculation:COV = average OD450 of negative controls x 2.1
Positive: OD450 of sample ≥ COV
Negative: OD450 of sample < COV
Invalid: If the average OD of negative controls is ≥ 0.1, the result is invalid. Repeat the test. If the problem persists, contact the local distributor.
Note: If the absorbance of negative controls is below 0.05, calculate it as 0.05. If the absorbance of negative controls is above 0.05, calculate it as its original value.
Performance Characteristics:Sensitivity: 2NCU/ml (per Biological Reference Reagents of Chinese Clinical Test Center), OD ≥ 1.500
Specificity: The average OD of 20 normal negative samples is ≤ 0.030
