Chemiluminiscence detection kit for 5 items of hepatitis B and preparation method thereof

A chemiluminescence detection and kit technology, applied in chemiluminescence/bioluminescence, analysis by making materials react chemically, measurement devices, etc., can solve problems such as unfavorable large-scale use, high cost, and economic burden, and achieve High degree of automation, saving manpower costs, and the effect of long-term testing platform

Active Publication Date: 2013-06-19
厦门市波生生物技术有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, most domestic quantitative tests for the 5 items of hepatitis B use imported test kits, which are expensive and costly. However, with a large population in my country, the

Method used

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  • Chemiluminiscence detection kit for 5 items of hepatitis B and preparation method thereof

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preparation example Construction

[0055] The preparation method of the five chemiluminescence quantitative detection kits for hepatitis B comprises the following steps:

[0056] 1. Preparation of Streptavidin Pre-coated Reaction Plate Preparation of Streptavidin Pre-coated Reaction Plate

[0057] Streptavidin was diluted with coating buffer. Take the reaction plate, and add the diluted streptavidin to the corresponding reaction plate, 150 μL per reaction plate. Place it at a constant temperature of 37°C for 2 hours for pre-coating. Shake off the coating solution, pat dry on clean absorbent paper, add 150 μL of sealing buffer to each reaction plate, and place at a constant temperature of 37°C for 2 hours. Remove the sealing solution, dry in a drying room at 37°C for 2 hours, and seal with a desiccant. The coating buffer is a Na2CO3-NaHCO3 buffer solution with a pH of 9.6 and 50 mmol / L, and the streptavidin dilution concentration is 8 μg / mL. The sealing buffer is 0.01mol / L TBS-T+(1%BSA, 0.2% gelatin). The r...

Embodiment 1

[0097] 1. Preparation of streptavidin pre-coated reaction plate

[0098] Streptavidin was diluted with coating buffer. Take the reaction plate, and add the diluted streptavidin to the corresponding reaction plate, 150 μL per reaction plate. Place it at a constant temperature of 37°C for 2 hours for pre-coating. Shake off the coating solution, pat dry on clean absorbent paper, add 150 μL of sealing buffer to each reaction plate, and place at a constant temperature of 37°C for 2 hours. Remove the sealing solution, dry in a drying room at 37°C for 2 hours, and seal with a desiccant. Described coating buffer is the Na of pH9.6, 50mmol / L 2 CO 3 -NaHCO 3 Buffer solution, the dilution concentration of streptavidin is 8 μg / mL. The sealing buffer is 0.01mol / L TBS-T+(1%BSA, 0.2% gelatin). The relative humidity of the drying room is less than 30%.

[0099] 2. Preparation of calibrator series

[0100] (1) Preparation of hepatitis B surface antigen calibrator

[0101] Dilute the ...

Embodiment 2

[0131] Similar to Example 1, the difference lies in the hepatitis B surface antibody in the specimen to be tested, and the sample, the hepatitis B surface antigen labeled with biotin and the hepatitis B surface antigen labeled with alkaline phosphatase are added to the solid phase of a microtube containing streptavidin. In the orifice plate, after reacting for 60 minutes, the hepatitis B surface antibody molecule specifically binds to the biotin-labeled hepatitis B surface antigen and the alkaline phosphatase-labeled hepatitis B surface antigen respectively, and the free components are washed away. Add luminescence substrate working solution, alkaline phosphatase substrate dephosphorylate, and emit 463nm visible light. The luminescence value RLU of each sample well was measured at the 15th minute. The RLU of the sample is positively correlated with the concentration of HBsAb in it. Sample results can be quantitatively calculated based on their RLU values.

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Abstract

The invention provides a chemiluminiscence detection kit for 5 items of hepatitis B and a preparation method thereof and relates to the kit for hepatitis B detection. The kit comprises a coated micropore reaction plate, a biotin marker, an alkaline phosphatase marker, a neutralization reagent, a calibrator, luminescent substrate liquid and a concentrated cleaning liquid. The preparation method comprises the following steps of: respectively preparing the streptavidin pre-coated reaction plate, the calibrator, the biotin marker, the alkaline phosphatase marker, the neutralization reagent, the luminescent substrate liquid and the concentrated cleaning liquid, and assembling for preparing the chemiluminiscence quantitative detection kit for 5 items of the hepatitis B. The prepared chemiluminiscence detection kit for 5 items of the hepatitis B has the advantages of being capable of sensitively detecting 5 items of the hepatitis B, also being capable of carrying out accurate quantitative determination on 5 items of the hepatitis B, being low in cost, simple in operation, accurate in results and environment-friendly without pollution and having long-term interests.

Description

technical field [0001] The invention relates to a kit for detecting hepatitis B, in particular to a five-item chemiluminescence detection kit for hepatitis B using enzyme-labeled biotin-streptavidin technology and a preparation method thereof. Background technique [0002] Hepatitis B (hepatitis B) is an infectious disease caused by hepatitis B virus (Hepatitis B Virus, HBV) infection that seriously endangers people's health. According to the WHO report, there are about 350 million HBV carriers in the world. my country is a high-incidence area for hepatitis B. According to statistics, the detection rate of hepatitis B surface antigen (HBsAg) in the Chinese population is 10.09%, and the number of HBsAg carriers is 120 million. Males are higher than females (10.3% vs. 7.29%), and rural areas are higher than urban (10.2% versus 7.9%), the total infection rate was as high as 35.5% to 61.1%, of which chronic hepatitis accounted for 56%. Every year, one million people die of liver...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/531G01N33/532G01N33/535G01N21/76
Inventor 陈巧钦严小莉蔡雅铨徐镜波杨家丽张长弓
Owner 厦门市波生生物技术有限公司
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