Preparation method and detection method of goat pox positive indirect hemagglutination diagnostic reagent

A diagnostic reagent, goat pox technology, applied in the field of preparation of goat pox positive indirect hemagglutination diagnostic reagent, can solve the problems of slow detection speed, low sensitivity, low specificity, etc., and achieve fast detection speed and high sensitivity , fast effect

Inactive Publication Date: 2011-11-23
GUIZHOU UNIV
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Problems solved by technology

The commonly used agar diffusion test detection method has the disadvantages of low sensitivity, slow detection speed, and low specificity.

Method used

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  • Preparation method and detection method of goat pox positive indirect hemagglutination diagnostic reagent
  • Preparation method and detection method of goat pox positive indirect hemagglutination diagnostic reagent

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Embodiment Construction

[0016] Embodiments of the present invention: the components of the goat pox positive indirect hemagglutination diagnostic reagent of the present invention are: pH 7.4 phosphate buffer, glutaraldehyde, sheep red blood cells, purified goat pox virus antigen, neonatal calf serum, NaN 3 .

[0017] Such as figure 1 Shown, the preparation method of goat pox forward indirect hemagglutination diagnostic reagent of the present invention, comprises the following steps:

[0018] First, aseptically collect venous blood from healthy sheep, add 10 times the amount of PBS, centrifuge at 2 000 rpm for 15 minutes, wash 5 times, and prepare 2.5% red blood cell suspension with 0.1M-pH 7.4-7.8 phosphate buffer ;

[0019] Second, mix the erythrocyte suspension prepared in step 1 with 1% glutaraldehyde at a volume ratio of 9:1, stir slowly at room temperature for 1 hour, centrifuge at 2000 rpm for 15 minutes, wash 5 times with phosphate buffer, and prepare Aldehyde red blood cells;

[0020] Th...

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Abstract

The invention discloses a goat pox positive indirect hemagglutination diagnostic reagent capable of detecting goat pox antibody and a preparation method and detection method thereof. The preparation method comprises the following steps: preparing red blood cell suspension by mixing glutaraldehyde and sheep red blood cells; mixing the red blood cell suspension with equivalent purified goat poxvirus, and stirring, centrifuging and washing; and preparing 1% sensitized red blood cells from PBS containing 1% of newly born calf serum to obtain the goat pox positive indirect hemagglutination diagnostic reagent. The optimized preparation conditions are that: the red blood cells are fixed by a glutaraldehyde single-hydroformylation method so that the concentration of sensitized red blood cells is 0.8-1.0%, and the antigen content is controlled between 20 ug.mL<-1> and 200 ug.mL<-1> so as to achieve relatively good sensitization effect; and the detection method is performed according to a conventional positive indirect hemagglutination test method, the reaction temperature is 20-25 DEG C, and the reaction time is 50 minutes. The method has relatively good specificity, sensitivity and repeatability, and can be applied to veterinarian clinical rapid detection of goat pox antibody.

Description

technical field [0001] The invention relates to a preparation method and a detection method of a goat pox positive indirect hemagglutination diagnostic reagent. Background technique [0002] Goat pox is an endemic highly contagious disease caused by goat poxvirus (GTPV). It can occur throughout the year, common in spring and autumn, regardless of gender, age, or variety. The World Organization for Animal Health (OIE) listed goat pox as a major infectious disease of category A, and in 1999, the Ministry of Agriculture of the People's Republic of China announced goat pox as a national first-class animal disease. In recent years, more than 20 provinces and cities in my country have successively reported the occurrence of goat pox epidemic, which has caused great economic losses and posed a serious threat to the booming goat breeding industry in my country. The commonly used detection method of agar diffusion test has the disadvantages of low sensitivity, slow detection speed ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/569
Inventor 刘忠伟周碧君程振涛文明王开功嵇辛勤岳筠汪德生
Owner GUIZHOU UNIV
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