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Reagent kit for screening infectious diseases and quantitating hepatitis B surface antigen and applications

A hepatitis B surface antigen and kit technology, applied in the field of medical detection, can solve the problems of inability to quantitatively detect hepatitis B surface antigen, inconvenient, fast, and incapable of quantitative detection, and reduce the detection cost, without washing, and with high accuracy. Effect

Pending Publication Date: 2019-09-24
北京协和洛克生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The joint detection products currently on the market include the colloidal gold method for qualitative joint detection of infectious diseases, and none of them can perform quantitative detection of hepatitis B surface antigen. Antigen to determine whether the patient is infected and the stage of infection, and accurately locate the treatment group and choose a treatment plan
It is necessary to do qualitative detection first and then quantitative detection, which is not convenient and fast enough, resulting in long detection time, low efficiency, high cost, and waste of manpower and material resources

Method used

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  • Reagent kit for screening infectious diseases and quantitating hepatitis B surface antigen and applications
  • Reagent kit for screening infectious diseases and quantitating hepatitis B surface antigen and applications
  • Reagent kit for screening infectious diseases and quantitating hepatitis B surface antigen and applications

Examples

Experimental program
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Effect test

Embodiment 1

[0087] This example is used to illustrate the preparation methods of the kit standard, quality control, sample buffer and positive control of the present invention.

[0088] The preparation of the concentration gradient standard substance and quality control substance of sAg: with the 10mM sodium phosphate buffer solution (PH7.4) that contains 2% neonatal calf serum, 0.05% Proclin300, 5% bovine serum albumin, 0.10% Triton X-100 The purchased sAg antigen 4.4 mg / ml was formulated into concentration gradient standards with marked concentrations of 0, 50, 150, 500, 1500, 5000 ng / ml and quality controls of 80 and 3000 ng / ml; then made into lyophilized powder, 4 Store at ℃;

[0089] Preparation of sample buffer: add 1% bovine serum albumin and 0.05% Proclin 300 respectively to 100 mM phosphate buffer (pH 7.4), and mix well.

[0090] The positive controls of hepatitis C virus antibody, HIV antibody and Treponema pallidum antibody were prepared by extracting goat serum and purifying ...

Embodiment 2

[0093] This example is used to illustrate the preparation of the hepatitis B virus surface antigen qualitative single item kit.

[0094] 1. Preparation of Hepatitis B surface antibody S11 coated Luminex No. 48 magnetic microspheres.

[0095] Principle: Use EDC to activate the carboxyl groups on the surface of the magnetic microspheres, and couple with the amino groups of the S11 antibody, so that the S11# antibody is coated on the magnetic microspheres.

[0096] method:

[0097] 1. Thoroughly mix Luminex No. 48 magnetic microspheres, and take out 0.2mL 1.25×10 7 pieces / ml.

[0098] 2. Use a magnetic stand for 60 seconds to separate the magnetic microspheres, and carefully aspirate the supernatant. 3. Add 0.1mL deionized water, shake for 60 seconds, sonicate for 20 seconds, separate on a magnetic stand for 60 seconds, suck out the supernatant, and repeat 2 times. 4. Add 0.08mL 50mm sodium dihydrogen phosphate pH4.0, shake for 20 seconds.

[0099] 5. Add 0.01 mL each of 50 ...

Embodiment 3

[0114] This example is used to illustrate the preparation of HIVP24 antigen single reagent.

[0115] 1. Preparation of P24 antibody-coated Luminex No. 72 magnetic microspheres.

[0116] Principle: Use EDC to activate the carboxyl groups on the surface of the magnetic microspheres, and couple with the amino groups of the HIVP24 antibody, so that the P24 antibody is coated on the magnetic microspheres.

[0117] method:

[0118] 1. Thoroughly mix Luminex No. 72 magnetic microspheres, and take out 0.2mL 1.25×10 7 pieces / ml.

[0119] 2. Use a magnetic stand for 60 seconds to separate the magnetic microspheres, and carefully aspirate the supernatant.

[0120] 3. Add 0.1mL deionized water, shake for 60 seconds, sonicate for 20 seconds, separate on a magnetic stand for 60 seconds, suck out the supernatant, and repeat 2 times. 4. Add 0.08mL 50mm sodium dihydrogen phosphate pH4.0, shake for 20 seconds.

[0121] 5. Add 0.01 mL each of 50 mg / mL EDC and NHS and mix well.

[0122] 6. ...

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Abstract

The invention provides a reagent kit for screening infectious diseases and quantitating a hepatitis B surface antigen and applications. The technology of the invention has the advantages of free combination, high throughput, high speed, low cost, high accuracy, god repeatability, wide linear range, no need for washing and simple operation. The introduction of liquid chip technology and products in clinical diagnosis can greatly improve the detection efficiency and reduce the detection cost. On a platform of the invention, the detection of human sAg concentration with high throughput, high speed, low cost, high accuracy and god repeatability can be realized, and quantitative experiments of HIV, HCV and TP can be carried out. Compared with conventional immunoassay, the invention aims to provide the high-throughput, rapid and accurate technical platform and reagent kit which can detect a plurality of infectious disease antigens and antibodies at the same time.

Description

technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a kit for infectious disease screening and hepatitis B surface antigen quantification and its application. Background technique [0002] Due to the spread of infectious diseases, it is necessary to detect infectious diseases before physical examination, blood donation, blood transfusion, and surgery. At present, the most widely transmitted and most harmful infectious diseases are AIDS, hepatitis B, hepatitis C and syphilis. Hepatitis B virus (HBV) surface antigen (HBsAg), hepatitis C virus (HCV) antibody (anti-HCV), human immunodeficiency virus (HIV) antibody (anti-HIV) and P24 antigen (HIVP24Ag), Treponema pallidum (TP) Antibody (anti-TP), that is, HBV, HCV, HIV and TP detection, is currently the main item for infectious disease screening. [0003] At present, the most commonly used method for detecting the above infectious diseases is immunoassay, which is based o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/571G01N33/68G01N33/577G01N33/533G01N33/543
CPCG01N33/56988G01N33/56983G01N33/571G01N33/6893G01N33/577G01N33/533G01N33/54326G01N2333/005G01N2446/90
Inventor 吴建榕娄金丽辛愿
Owner 北京协和洛克生物技术有限责任公司
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