Specific-antigen preparing method used for detecting schistosoma japonicum antibodies by indirect hemagglutination (IHA) method

A schistosomiasis and antigen technology, which is applied in the field of specific antigen preparation for Schistosoma japonicum antibody detection, can solve the problems of complex components, low specificity, and high cross-reactivity, and achieve high sensitivity, strong specificity, and reduced cross-reaction effects

Pending Publication Date: 2019-12-24
安徽省血吸虫病防治研究所
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Problems solved by technology

The mixed specific antigen based on the soluble antigen of schistosomiasis eggs developed by us is used as the antigen to make the indirect haemagglutination diagnostic reagent for schistosomiasis, which overcomes the shortcomings of the traditional single worm egg crude antigen with complex components and high cross-reactivity. Recombinant antigen, low specificity defect

Method used

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Embodiment 1

[0027] A kind of specific antigen preparation method that is used for indirect hemagglutination (IHA) method to detect Schistosoma japonicum antibody, comprises the following steps:

[0028] Step 1: Preparation of schistosomiasis-specific antigen:

[0029] Step 1, prepare the soluble adult worm antigen of Schistosoma japonicum; take the freeze-dried product of adult Schistosoma japonicum, add 50mmol / L sodium acetate buffer solution with pH5.6 at a concentration of 1%, and freeze-thaw repeatedly between -20°C and 37°C for 3 times , sonicate for 10 minutes, put it in the refrigerator overnight, centrifuge at 10,000 g at 4°C for 30 minutes, absorb the supernatant, filter through filter paper, and centrifuge again at 10,000 g at 4°C for 30 minutes. The sodium buffer solution was prepared to a final concentration of 1 mg / ml for later use.

[0030] Step 2. Prepare Schistosoma japonicum soluble cercariae antigen; take the freeze-dried product of Schistosoma japonicum cercariae, add ...

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Abstract

The invention provides a specific-antigen preparing method used for detecting schistosoma japonicum antibodies by an indirect hemagglutination (IHA) method. The method includes the following steps: step one, preparing schistosoma-specific antigens, wherein in a step 1, soluble adult antigens of schistosoma japonicum are prepared, in a step 2, soluble cercaria antigens of the schistosoma japonicumare prepared, in a step 3, soluble egg antigens of the schistosoma japonicum are prepared, in a step 4, the antigens are treated by sodium periodate, and in a step 5, the soluble adult antigens of theschistosoma japonicum, the soluble cercariae antigens of the schistosoma japonicum and the soluble egg antigens of the schistosoma japonicum after treating of the sodium periodate are proportionallymixed according to a volume ratio of 1:0.5:2, that is, specific antigens are obtained; and step two, preparing an indirect hemagglutination reagent by the specific antigens. According to the method, the crudely prepared soluble antigens of schistosoma japonicum adults, cercariae and eggs are proportioned and mixed after treating of the sodium periodate, the diagnostic reagent is prepared, and thereagent reduces cross-reaction with other diseases, and maintains original high sensitivity and specificity at the same time.

Description

technical field [0001] The invention relates to the technical field of schistosome antibody detection, in particular to a method for preparing a specific antigen used in the detection of schistosome antibody in an indirect hemagglutination reagent. Background technique [0002] Schistosomiasis is a major infectious disease that seriously endangers the health of the population and hinders economic and social development. The party and government of our country attach great importance to the prevention and control of schistosomiasis. After more than 60 years of hard work, the prevention and control of schistosomiasis in my country has achieved world-renowned achievements. However, in the process of controlling and eliminating schistosomiasis, we still have many technical barriers and obstacles. Especially with the in-depth prevention and control of schistosomiasis, the infection rate of schistosomiasis in endemic areas is getting lower and lower, and the infection degree is ge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/577G01N33/531
Inventor 汪天平王恩木章乐生张世清呼明闯
Owner 安徽省血吸虫病防治研究所
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