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In vitro detection method of hepatitis B virus C antibody

A hepatitis B virus, in vitro detection technology, applied in measurement devices, color/spectral property measurement, instruments, etc., can solve problems such as the inability to meet the needs of large-scale in vitro detection

Inactive Publication Date: 2011-05-04
SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These technologies can only meet the needs of scientific research at present, and cannot meet the needs of large-scale in vitro testing in medical and clinical observation.

Method used

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  • In vitro detection method of hepatitis B virus C antibody
  • In vitro detection method of hepatitis B virus C antibody
  • In vitro detection method of hepatitis B virus C antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1Protein A is coated on microtiter plate

[0067]Select an untreated polystyrene microtiter plate, add 0.9μg / ml Protein A dissolved in carbonate buffer, pH9.2-9.8, 100μl / well, overnight at 4°C. The coating solution was removed, and 200 μl of 1% (w / v) BSA solution (PBS, pH 7.4) was added to each well for blocking, at 37° C. for 1-2 hours.

Embodiment 2

[0068] Embodiment 2 Serum sample detection method

[0069] Wash the microtiter plate 1-2 times with PBST (PBS buffer containing 0.05% Tween-20), and pat dry each time.

[0070] After adding 90 μl of PBS to each well, add 10 μl of serum to be tested into each well, mix well and incubate at 37° C. for 60 minutes.

[0071] After the reaction solution was removed and patted dry, the plate was washed 4 times with PBST, each time patted dry.

[0072] Add 40 μl of recombinant HBcAg (SEQ ID NO: 1, provided by Shanghai Rongsheng Biopharmaceutical Co., Ltd.) (diluted 1:2000 with PBS) and 40 μl of HRP-labeled HBcAg antibody (diluted 1:2000 with PBS) into each well.

[0073] Add 50 μl each of substrate A and B solution to each well, mix well, incubate at 37°C for 15 minutes, then add stop solution, and measure with a 450nm microplate reader.

[0074] Substrate A solution preparation:

[0075] Weigh 17.9g of citric acid and Na 2 HPO 4 12 2 O 4.67g was dissolved in 400ml deionized wat...

Embodiment 3

[0081] Embodiment 3HBcAb detects parallel control

[0082] The currently used HBcAb competition ELISA detection kit (Shanghai Rongsheng Biopharmaceutical Co., Ltd.) was used for comparison.

[0083] A total of 350 specimens were used in this experiment, among which,

[0084] Da San Yang (positive for HBsAg, HBeAg and HBcAb): 110 cases;

[0085] Small Sanyang (positive for HBsAg, HBeAb and HBcAb): 100 cases;

[0086] Healthy human serum (from blood bank): 140 cases.

[0087] The test results are shown in Table 1.

[0088] Table 1 HBcAb detection parallel control

[0089]

[0090] It can be obtained from Table 1:

[0091] Positive consistency rate: 203 / 203=100%;

[0092] Negative consistency rate: 140 / 147=95.2%;

[0093] Overall agreement rate: 343 / 350=98%.

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Abstract

The invention relates to an in vitro detection method of a hepatitis B virus C antibody. A hepatitis B virus C antigen with more than two antibody recognition epitopes is used for simultaneously combining an antibody to be detected with an antibody combined with a label, and the antibody to be detected can be determined by directly detecting the label or detecting after a reaction substrate is added. The detection result is consistent to the recognition habit of human and is more convenient to operate and judge. The method also increases the sensitivity and the repeatability of detection and enhances detection specificity.

Description

technical field [0001] The invention relates to a detection method of hepatitis B virus antibody, in particular to an in vitro detection method of hepatitis B virus C antibody. Using this method to detect the hepatitis B virus C antibody can make the detection more accurate and the result more intuitive. Background technique [0002] Hepatitis B (viral) is a worldwide disease caused by hepatitis B virus (HBV). The incidence rate is high in developing countries. According to statistics, there are more than 280 million asymptomatic hepatitis B virus carriers in the world. my country accounts for about 93 million people, most of them are asymptomatic, but 1 / 3 of them will have clinical manifestations of liver damage. [0003] At present, there are three ways to diagnose hepatitis B, such as: detecting the antigen of the virus, detecting the antibody in the body, and detecting the DNA of the virus. Clinically, five markers are usually used for judgment, such as: hepatitis B s...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/577G01N21/31
Inventor 朱绍荣
Owner SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
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