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Method for detecting schistosome circulating antigen and enzyme-linked immune kit thereof

An enzyme-linked immunosorbent reagent and technology of schistosomiasis, applied in the field of bioengineering, can solve the problems of low sensitivity and specificity, and achieve the effect of high sensitivity, strong specificity and easy operation

Inactive Publication Date: 2011-02-09
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 object of the present invention is to provide a method for detecting schistosomiasis circulating antigen and its ELISA kit, the detection method of this schistosomiasis circulating antigen and its ELISA kit should solve the problem of detecting schistosomiasis circulating antigen in the prior art Technical problems of low sensitivity and low specificity of the method

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  • Method for detecting schistosome circulating antigen and enzyme-linked immune kit thereof
  • Method for detecting schistosome circulating antigen and enzyme-linked immune kit thereof
  • Method for detecting schistosome circulating antigen and enzyme-linked immune kit thereof

Examples

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Effect test

Embodiment 1

[0037] Example 1 The preparation method of anti-Schistosoma japonicum soluble egg antigen polyclonal antibody IgY is as follows:

[0038] 1. Preparation of soluble antigen of Schistosoma japonicum: 7 New Zealand rabbits each infected with 1500 cercariae of Schistosoma japonicum (from China Center for Disease Control and Prevention, Parasitic Disease Prevention and Control Institute) were dissected according to the conventional method after 42 days, and separated and collected from the liver eggs and freeze-dried. Take the dry eggs and weigh them, dissolve them in 0.9% physiological saline at a ratio of 1%, grind them, and place them in a cold soak at 4°C, during which they shake 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, meas...

Embodiment 2

[0041] The preparation method of embodiment 2 anti-schistosome SEA monoclonal antibody is as follows:

[0042] (1) Animal immunity

[0043] ①Preparation of antigen: 7 New Zealand rabbits each infected with 1500 cercariae of Schistosoma japonicum (from the Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention) were dissected according to conventional methods after 42 days, and eggs were collected from the liver and freeze-dried . Take the dry eggs and weigh them, dissolve them in 0.9% physiological saline at a ratio of 1%, grind them, and place them in a cold soak at 4°C, during which they shake 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 me...

Embodiment 3

[0071] Embodiment 3 A kind of method for detecting schistosomiasis circulating antigen

[0072] Such as figure 1 As shown, a method for detecting schistosome circulating antigens of the present invention comprises the steps:

[0073] (1) Select the polyclonal antibody IgY against the soluble antigen of Schistosoma japonicum to be used for coating the solid phase carrier (1), and the monoclonal antibody against the soluble antigen of Schistosoma japonicum egg is used to prepare the enzyme conjugate;

[0074] (2) Put human or mouse or rabbit serum to be tested (2) on the solid-phase carrier (1) coated with polyclonal antibody IgY anti-schistosomiasis egg soluble antigen, after incubation and washing, add enzyme Label the monoclonal antibody (3) against the soluble antigen of Schistosoma japonicum eggs, after incubation and washing, the polyclonal antibody IgY-circulating antigen-enzyme-labeled anti-Schistosomiasis SEA monoclonal antibody sandwich complex (4 ), positive samples...

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Abstract

The invention relates to a method for detecting a schistosome circulating antigen, which comprises the following steps of: preparing a schistosome soluble egg antigen (SEA) resistant specific IgY polyclonal antibody; preparing an enzyme-labeled schistosome SEA resistant monoclonal antibody by adopting a hybridoma technology; selecting a polyclonal antibody for coating a solid-phase carrier; selecting a schistosome SEA resistant monoclonal antibody for preparing an enzyme composition; forming a schistosome SEA resistant specific IgY polyclonal antibody-circulating antigen-enzyme-labeled schistosome SEA resistant monoclonal antibody sandwich complex; and adding a substrate for developing and indicating. The invention also provides an enzyme-linked immune kit for detecting the schistosome circulating antigen. The method for detecting schistosome circulating antigen has the advantages of high sensitivity, high specificity and high repeatability and is respectively suitable for the detection of mass specimens and curative effect assessment.

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 method for detecting circulating antigens of schistosomiasis and an enzyme-linked immunosorbent kit thereof. Background technique [0002] 1. The importance of immunodiagnosis in the prevention and treatment of schistosomiasis japonicum [0003] Schistosomiasis is a zoonotic parasitic disease widely prevalent in tropical and subtropical regions. About 200 million people in 76 countries and regions around the world are infected, and 600 million people are threatened by infection. my country is an endemic area of ​​schistosomiasis japonicum, which seriously affected people's normal production and life in the early days of the founding of the People's Republic of China. After more than 50 years of prevention and control, remarkable achievements have been made. By 2003, the total number of schistosomiasis patients in Chin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/52G01N33/577G01N33/569
Inventor 陈家旭郭俭蔡玉春陈木新陈韶红田利光
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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