Specifications

1.Double antibody sandwich EIA to detect Hepatitis B surface antigen
3.Used for diagnosis of HBV infection
4.analytical test

 

Certificate

GMP

SFDA

ISO13485

ISO9001:2008

SPECIMEN COLLECTION AND PREPARATION

 

Serum should be prepared from a whole blood specimen obtained by acceptable medical techniques. Either serum or plasma can be used in this test.  Remove serum or plasma from the clot or blood cells as soon as possible to avoid hemolysis.  Specimen with extensive particulate should be clarified by centrifugation prior to use.  Specimen frozen at -20°C or colder may be used.  Avoid repeated freeze thaw.

 

 

1. 1.        For in-vitro diagnostic use only.
2. 2.        Do not use kit beyond expiration date.
3. 3.        Do not mix components from kits with different lot number.
4. 4.        Avoid microbial contamination of reagents.
5. 5.        Do not pipet reagent by mouth and no smoking or eating while performing assays.
6. 6.        Wear gloves during the whole process and avoid reagents or specimen spilling-out.
7. 7.        Wipe up the spills using 5% hypochlorite solution.
8. 8.        Decontaminate all liquids or solid wastes before deposing.

 

STORAGE OF TEST KITS

 

Unopened test kits should be stored at 2-8°C. DO NOT FREEZE KIT COMPONNETS.  The microtiter plate should be kept in a sealed bag to minimize exposure to damp air. Use up the reagents as soon as possible after the kit is unpacked.

 

WORKING WASH BUFFER

 

Dilute the 20X wash buffer concentrate with deionized or distilled water 1:20. For example, 5 ml of wash buffer concentrate should be diluted to a total volume of 100 mL with deionized or distilled water.

 

ASSAY PROCEDURE:

  

It is strongly advised to analyze each specimen and controls in duplicate. All the reagents should equilibrate to room temperature before use.

1. 1.        Dispense one drop (100μl) of Positive Control as well as Negative Control in duplicate into respective wells. Set one blank well as background control, and 100ul of serum or plasma samples into respective test wells.
2. 2.        Place the microtiter plates into a humidified box, and incubate at 37°C for 60 minutes.
3. 3.        Add one drop (50μl) of Enzyme Conjugate to each well. Mix it gently by swirling the microtiter plate on flat bench for 1 minute. Do not add Enzyme Conjugate to the blank well.
4. 4.        Place the microtiter plates into a humidified box, and incubate at 37°C for 30 minutes.
5. 5.        Wash each well 5 times by filling each well with diluted wash buffer, then inverting the plate vigorously to get all water out and blocking the rim of wells on absorbent paper for a few seconds.
6. 6.        Add one drop (50μl) of Substrate Solution A (HRP-substrate) to each well, then add one drop (50μl) of Substrate Solution B (TMB) to each well. Mix gently and incubate at 37°C for 30minutes.
7. 7.        Add 1 drop (50μl) of Stop Solution to each well to stop the color reaction. Read the OD value at 450 nm/630 nm with dual filter plate reader. It is option to read the OD value at 450 nm with single filter plate reader. (using the OD value of the blank well to correct all the OD reading from all wells)

 

 

 

INTERPRETATION OF RESULTS

 

1. A run is valid if:

1)   The full complement of Blanks, Positive and Negative Controls must be included in each assay.

2)   Negative Control values must have an absorbance and ≤0.10 after subtracting the Blank.

3)   Positive Control value must have absorbance ≥1.00 after subtracting the Blank.

     

2. Calculation of Cut off Value (COV)

Mean of the Negative Controls (NCx)* 2.1

 

If the OD value of the negative control is less than 0.05, it should be reported as 0.05. If it is more than 0.05, it should be reported as the actual OD value measured.

Positive OD reading:    ≥Cut-off value

Negative OD reading:  < Cut-off value

 

TESTPERFORMANCEANDEXPECTEDRESULTS

 

This assay was standardized against Reference Standards provided from the Reference Laboratory for Immunology Product under the Ministry of Health, China.

 

Clinical Specificity: The clinical specificity of this assay was determinate by a panel of samples obtained from 2500 healthy blood donors and 300 undiagnosed hospitalized patients. The repeatedly reactive samples and samples confirmed positive with the reference test were not included in the calculation of specificity.