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In vitro detection method and application of antibody

An in vitro detection and antibody technology, applied in measuring devices, color/spectral characteristic measurement, instruments, etc., can solve the problems that cannot 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 and application of antibody
  • In vitro detection method and application of antibody
  • In vitro detection method and application of antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1Protein A is coated on microtiter plate

[0068] 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, overnight at 4°C or 1-2 hours at 37°C.

Embodiment 2

[0069] Embodiment 2 Serum sample detection method

[0070] Wash the microtiter plate 1-2 times with PBST (PBS buffer containing 0.05% Tween-20), and pat dry each time.

[0071] After adding 50 μl of PBS to each well, add 50 μl of serum to be tested into each well, mix well and incubate at 37° C. for 60 minutes.

[0072] After the reaction solution was removed and patted dry, the plate was washed 4 times with PBST, each time patted dry.

[0073] Add 50 μl recombinant HBeAg (SEQ ID NO: 2, provided by Shanghai Rongsheng Bio-Pharmaceutical Co., Ltd.) (diluted 1:6000 with PBS) and 50 μl HBeAg antibody labeled with HRP (diluted 1:4000 with PBS) to each well.

[0074] 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.

[0075] Substrate A solution preparation:

[0076] Weigh 17.9g of citric acid and Na 2 HPO 4 12 2 O 4.67g was dissolved in 400ml deionized water...

Embodiment 3

[0082] Embodiment 3HBeAb detects parallel control

[0083] The currently used HBeAb competition ELISA detection kit (Shanghai Rongsheng Biopharmaceutical Co., Ltd.) was used for comparison.

[0084] A total of 350 specimens were used in this experiment, among which,

[0085] Da San Yang (positive for HBsAg, HBeAg and HBcAb): 110 cases;

[0086] Small Sanyang (positive for HBsAg, HBeAb and HBcAb): 100 cases;

[0087] Healthy human serum (from blood bank): 140 cases,

[0088] The test results are shown in Table 1.

[0089] Table 1 HBeAb detection parallel control

[0090]

[0091] It can be obtained from Table 1:

[0092] Positive consistency rate: 100 / 100=100%;

[0093] Negative consistency rate: 235 / 250=94%;

[0094] The overall agreement rate: 335 / 350=95.7%.

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Abstract

The invention relates to an in vitro detection method of an antibody. An antibody to be detected can be determined by using an antigen with more than two antibody recognition epitopes to simultaneously combine with the antibody to be detected and the antibody with a marker to directly detect the marker or detect the marker after adding a reaction substrate. The detection result is consistent with the cognition habit of people, the in vitro detection method is more convenient to operate and judge. With the method, the sensitivity and the repeatability of the detection are improved and the detection specificity is enhanced.

Description

technical field [0001] The invention relates to an antibody detection method, in particular to an antibody detection method in vitro. After the method is applied to detect the hepatitis B virus E antibody, the detection result can be more intuitive and accurate, and the hepatitis B virus E antibody in the serum can also be quantified. 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 j...

Claims

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Application Information

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