Kit and method for detecting circulating antigen of oriental schistosomiasis

A schistosomiasis and kit technology, applied in the field of bioengineering, can solve the problems of low specificity and low sensitivity, and achieve the effects of strong specificity, high sensitivity and easy operation.

Inactive Publication Date: 2011-04-27
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to provide a test kit and method for detecting schistosomiasis circulating antigen, said test kit and method for detecting schistoso

Method used

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  • Kit and method for detecting circulating antigen of oriental schistosomiasis
  • Kit and method for detecting circulating antigen of oriental schistosomiasis
  • Kit and method for detecting circulating antigen of oriental schistosomiasis

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1 The preparation method of biotin-labeled anti-Schistosoma japonicum soluble egg antigen polyclonal antibody IgG is as follows:

[0042] 1. Preparation of soluble egg antigen (SEA) of Schistosoma japonicum: Seven New Zealand rabbits infected with 1500 cercariae of Schistosoma japonicum were dissected according to the conventional method after 42 days, and the eggs were collected from the liver and freeze-dried. Take the dry eggs and weigh them, dissolve them in 0.9% normal saline at a ratio of 1%, place them in a cold soak at 4°C for 4 days, and shake them twice a day for 2 minutes each time. After 4 days, ultrasonic pulverization was performed in an ice bath, 3 minutes each time, with a 3-minute interval, 3 times in total. Then cold soak for 2 days as above, centrifuge at 10,000 rpm at 4°C for 30 minutes, take the supernatant, collect and aliquot, measure the concentration of antigenic protein by Braford method, and store at -20°C for later use.

[0043] 2. A...

Embodiment 2

[0046] Example 2 The preparation method of the electrochemiluminescent agent-labeled anti-Schistosoma japonicum SEA-specific monoclonal antibody is as follows:

[0047] 1. Antigen immunization: SEA antigen was prepared according to the method in Example 1. Take 75 μl of SEA antigen with a protein concentration of 4 mg / ml, mix it with an equal volume of Freund's complete adjuvant, fully emulsify, and inject 5 BALB / C mice intraperitoneally, 0.5 ml each. Four weeks later, the same dose of SEA antigen was fully emulsified with an equal volume of Freund's incomplete adjuvant to boost the immunization once, and the injection dose was the same as above. Two weeks later, the same amount of antigen was directly used to boost the immunization once, and the injection dose was the same as the first time. After 2 weeks, the serum antibody level was measured, and 2 mice with high antibody titer were selected, and 50 μg / 0.2ml of each mouse was injected with the antigen through the tail vein...

Embodiment 3

[0058] Embodiment 3 A method for detecting circulating antigens of Schistosoma japonicum comprises the following steps:

[0059] (1) Selecting anti-Schistosoma japonicum SEA polyclonal antibody IgG for preparing biotin-labeled conjugate 2, and anti-Schistosoma japonicum SEA-specific monoclonal antibody for preparing electrochemiluminescent agent-labeled conjugate 4;

[0060] (2) Streptavidin magnetic beads 1 purchased from Norway Dynal company and anti-Schistosoma japonicum SEA polyclonal antibody IgG biotin-labeled conjugate 2 were added to the reaction tube, incubated for reaction and washed, and serum test sample 3 was added , after incubation and washing, add an electrochemiluminescence agent to label anti-Schistosoma japonicum SEA monoclonal antibody 4, after incubation and washing, form streptavidin magnetic beads-biotin-labeled anti-Schistosoma japonicum SEA polyclonal antibody IgG-circulation Antigen-electrochemiluminescent agent labeled anti-Schistosoma japonicum SEA ...

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Abstract

The invention provides a kit for detecting the circulating antigen of oriental schistosomiasis, comprising streptavidin-coated magnetic beads, an oriental schistosomiasis resistant soluble egg antigen polyclonal antibody marked by biotin, an oriental schistosomiasis resistant soluble egg antigen specificity monoclonal antibody marked by an electrochemiluminescence reagent, a cleaning solution, a sample diluent, schistosomiasis positive serum, schistosomiasis negative serum and a co-reaction reagent. The invention further provides a method for detecting the circulating antigen of oriental schistosomiasis. In the invention, the kit for detecting the circulating antigen of oriental schistosomiasis is simple, convenient and fast in operation, is suitable for the clinic diagnosis, the curative effect check and the epidemiological investigation at the departments of hospital clinical laboratory, disease prevent and control center, community healthy center and the like, and has very high use value; and the double-antibody sandwich method for detecting the circulating antigen of oriental schistosomiasis has high sensitivity, strong specificity and good repeatability, and is suitable for the detection of batches of specimens and the curative effect check.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a rapid diagnosis method for schistosomiasis, in particular to a kit and method for detecting circulating antigens of Schistosoma japonicum. Background technique [0002] 1. The importance of immunodiagnosis in the prevention and treatment of schistosomiasis japonicum [0003] Schistosomiasis is a zoonotic parasitic disease that seriously endangers human health. The number of infected people in the world is about 200 million, and 600 million people are threatened by infection. my country is an endemic area of ​​schistosomiasis. After more than 50 years of prevention and control, it has achieved world-renowned achievements. By 2003, the number of schistosomiasis patients in China had decreased by 92.74% compared with the early days of the founding of the People's Republic of China. Five provinces (autonomous regions and municipalities) including Guangdong, Shanghai, Fujian, Guangxi,...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/76
Inventor 陈家旭郭俭蔡玉春陈木新陈韶红李浩张永年陈海宁储言红周卉田利光
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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