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PRINCIPLE OF THE ASSAY for  HBeAg

The Bioneovan HBsAb ELISA kit uses antigen “sandwich” ELISA method where polystyrene microwell strips are pre-coated with recombinant HBsAg. Serum or plasma sample is added to the microwells together with a second HBsAg conjugated to Horseradish Peroxidase (HRP-Conjugate). When antibodies to HBsAg are present in the sample, the pre-coated and conjugated antigens will be bound to the two variable domains of the antibody ,the specific immunocomplex formed is captured on the solid phase during incubation. Non-reactive antibodies and unbound HRP-Conjugates are removed with the wash buffer. The enzyme substrate, tetramethylbenzidine (TMB), is added upon incubation the substrate will be hydrolyzed by the bound enzyme and a blue or blue-green colour develops in wells containing anti-HBs. The enzyme reaction is stopped by the addition of sulphuric acid. The intensity of colour developed is read spectrophotometrically at 450nm (450 nm/630 nm) and is proportional to the amount of antibodies present in the specimen.No color or very pale blue color indicates a negative reaction.

SPECIMEN COLLECTION AND PREPARATION  for  HBeAg

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℃ or colder may be used. Avoid repeated freeze thaw.

ASSAY PROCEDURE  for HBeAg  : 

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

1. For each test, set one blank well as background control, two positive and two negative controls.

2. Add 50μl positive control, Negative control, and Specimen into their respective wells. Neither samples nor HRP-Conjugate should be added into the Blank well.

3. Add 50 μl Enzyme Conjugant to each well except the Blank. Mix it gently by swirling the microtiter plate on flat bench for 1 minute. Do not add Enzyme Conjugate to the blank well.

4. Cover wells with seal paper, Place the microtiter plates into a humidified box, and incubate at 37℃ for 30 minutes.

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. Add one drop (50 ul) of Substrate Solution A (HRP-substrate) to each well, then add one drop (50 ul) of Substrate Solution B (TMB) to each well. Mix gently and incubate at 37℃ for 15 minutes. .

7. Add 1 drop (50 ul) 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  for  HBeAg

EIA Reader at 450 nm (using the OD value of the blank well to correct all the OD reading from all wells or at 450 nm/630 nm with dual filter plate reader.

Cut-off Calculations:

Take average OD values of Negative control and times 2.1:

2.1 ×NC x = Cut-off.

If the OD value of the negative control is less than 0.07, it should be reported as 0.07. If it is more than 0.07, it should be reported as the actual OD value measured.

Positive OD reading: ≥Cut-off value

Negative OD reading: < Cut-off value

We will send free sample can represent our bulk cargo for our client’s reference. 

1. For in vitro diagnostic test use only 
2. Do not use test kit beyond expiry date 
3. The test device should not be reused

Q:Can I get a sample before bulk order? How can I get it?

A:Sample is free, but freight collect.

You could prepay the freight cost by PayPal or Western union, we will send out sample ASAP once getting your payment.

Q:How about supply ability?

A:100,000 pcs per week for cassette, 500,000pcs per week for strip.

Q: How to ensure the cargo quality?

A: We will supply bulk sample before shipping. They can represent the cargo quality.

Q: Why choose Bioneovan?

A: We are the professional factory not trading company, so the price is quite competitive in the market.

We have ISO13485 & CFDA & GMP certificate. Products quality can be guaranteed.

Q: What’s the payment method?

A: T/T: Accept USD.

Western Union/Money Gram/PayPal: Quickly to the account, Priority in delivery

Behalf Paying: Your Chinese friends or your Chinese agent can pay for in RMB.

Q: How to contact us?

Bioneovan Co.,Ltd.

Name: Snow Liu

Whats App:+86 15630062340

Tel: +86-10-69255832-8006

Website: www.bioneovan.com    http://bioneovan.en.alibaba.com/

Snow

Email: snow @bioneovan.net

Tel: +86-10-69255832-8006

Whats app:+86 15630062340

QQ: 3523788440

Address: Keyuan Road 18 Daxing Industry Zone Beijing, China

Please contact me if necesarry, it's my pleasure to help you.