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HCV core antigen detection kit based on magnetic micro-particle chemiluminescence method

A chemiluminescence method and a detection kit technology, applied in chemiluminescence/bioluminescence, analysis through chemical reactions of materials, measurement devices, etc., can solve the problems of long window period, difficult promotion, low sensitivity, etc., and achieve easy The effects of automated operation, expanded detection range, and shortened window period

Inactive Publication Date: 2015-02-18
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the above prior art anti-HCV ELISA reagents have false positives, long window period, low sensitivity and RT-PCR detection of HCV RNA requires high technology and equipment, time-consuming and laborious, difficult to promote in routine work or grassroots laboratories and other shortcomings

Method used

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  • HCV core antigen detection kit based on magnetic micro-particle chemiluminescence method
  • HCV core antigen detection kit based on magnetic micro-particle chemiluminescence method
  • HCV core antigen detection kit based on magnetic micro-particle chemiluminescence method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The composition and preparation method of the kit

[0029] 1. Reagent R1: Add 0.1 g of magnetic beads coated with anti-HCV-cAg antibody purchased from Dynal to 100 ml of 0.1M PBS buffer with pH=7.6 and mix well.

[0030] 2. The preparation method of HRP-labeled anti-HCV-cAg antibody in reagent R2 is as follows:

[0031] (1) Weigh 25 mg of HRP and dissolve it in 1.25% glutaraldehyde solution, and let it stand overnight at room temperature.

[0032] (2) The reacted enzyme solution was eluted with a Sephadex G-25 chromatographic column with physiological saline. The flow rate was controlled at 1ml / min, and the brown effluent was collected. If the volume is greater than 5ml, concentrate to 5ml with PEG. Place in a 25ml small beaker and stir slowly.

[0033] (3) Dilute 12.5mg of the antibody to be labeled to 5ml with physiological saline, and add it dropwise to the enzyme solution while stirring.

[0034] (4) Add 0.25ml of 1M pH9.5 carbonic acid buffer and continue to s...

Embodiment 2

[0046] Sensitivity and linear range experiments

[0047] Through experimental detection and analysis, the determination of the cut off value of the reagent: the average RLU value of the negative reference product detection × 2.1 (if the average RLU value of the negative reference product detection is ≤ 60, calculate as 60; when RLU > 60, calculate according to the actual Calculate the average RLU value of the negative control × 2.1). The low-value samples were detected, and the luminescence values ​​of the negative reference product and normal human serum were compared at the same time, as shown in Table 1. By comparison, when the low-value sample concentration is 0.5ng / mL, it is still detected as positive, indicating that the sensitivity of the reagent in Example 1 can reach 0.5ng / mL, which has good sensitivity.

[0048] Table 1 Sensitivity determination

[0049] Test items Luminescence value (RLU) Result judgment HCV-cAg 0.5ng / mL 331 + 20 normal h...

Embodiment 3

[0052]The detection kit for hepatitis C core antigen by magnetic particle chemiluminescence method of the present invention has a good correlation with the result obtained by real-time fluorescent quantitative PCR.

[0053] The specific method is as follows: 40 cases of clinical samples are detected simultaneously with the kit obtained in Example 1 and the hepatitis C virus nucleic acid detection kit (PCR-fluorescent probe method) produced by Sun Yat-sen University Daan Gene Co., Ltd., and the results are very good. good correlation, specific results such as figure 2 shown.

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Abstract

The invention relates to an HCV core antigen detection kit based on a magnetic micro-particle chemiluminescence method, which belongs to the technical field of clinical in-vitro detection. Total or free HCV core antigens in a serum sample are qualitatively or quantitatively detected by virtue of a double-antibody sandwich method. The HCV core antigen detection kit mainly comprises a reagent R1, a reagent R2, a bottle of positive reference, a bottle of negative reference, a luminous substrate A, a luminous substrate B and a solid washing solution. The kit is high in sensitivity and stability, is simple, convenient and quick to operate, is easy to automatically operate, and can well meet domestic clinical using requirements.

Description

technical field [0001] The invention relates to a detection kit for hepatitis C core antigen by magnetic particle chemiluminescence method, which qualitatively or quantitatively detects total or free HCV core antigen in a serum sample by a double-antibody sandwich method, and belongs to the technical field of clinical in vitro detection. Background technique [0002] Type C viral hepatitis , referred to as hepatitis C, hepatitis C, is a viral hepatitis caused by hepatitis C virus (HCV) infection, mainly through blood transfusion According to the statistics of the World Health Organization, the global HCV infection rate is about 3%, it is estimated that about 180 million people are infected with HCV, and there are about 35,000 new cases of hepatitis C every year. Hepatitis C is a global epidemic, which can lead to chronic inflammation, necrosis and fibrosis of the liver, and some patients can develop liver cirrhosis Even hepatocellular carcinoma (HCC). Mortality associat...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/553G01N21/76
CPCG01N33/5767G01N21/76G01N33/553
Inventor 甘宜梧李志明谭柏清谢清华王绮肖慧朱玉娥
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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