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Mycoplasma capricolum subsp. pneumonia antigen of goats and preparation method thereof

The technology of mycoplasma and goat is applied in the field of preparation of antigen of Mycoplasma capricolum capitis subspecies and indirect hemagglutination diagnostic reagents for detecting antibodies of Mycoplasma capricolum capitis subspecies, which can solve the lack of epidemiological information of Mycoplasma capricolum capitis subspecies and epidemiological information. It is easy to store for a long time, low price, and high blood coagulation titer.

Active Publication Date: 2010-05-26
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the actual prevalence of this disease in my country and accurate epidemiological information are still relatively scarce. The main reason for this phenomenon is that my country currently lacks relevant technologies for the effective detection of Mycoplasma capricosum subsp.

Method used

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  • Mycoplasma capricolum subsp. pneumonia antigen of goats and preparation method thereof
  • Mycoplasma capricolum subsp. pneumonia antigen of goats and preparation method thereof
  • Mycoplasma capricolum subsp. pneumonia antigen of goats and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0019] 1 culture mycoplasma

[0020] 1.1 Culture medium preparation

[0021] Prepare 1200ml of modified KM2 medium. Mix 420ml of 1.7% hydrolyzed milk protein Hank's buffer, 24ml of 25% yeast infusion, 12ml of 1% thallium acetate solution, and 12ml of 0.4% phenol red. Membrane filtration sterilized minimum element medium (MEM) 600ml, healthy horse serum 120ml, 20,000 IU penicillin 12ml, adjusted the pH value to 7.4 with 1mol / L sodium hydroxide solution, and stored at 8°C for later use.

[0022] 1.2 Mycoplasma culture

[0023] Inoculate 5ml of the improved KM2 culture medium with Mycoplasma capricosum subspecies goat pneumonia, place 37 ℃ and cultivate for 6 days, the color of the culture medium turns yellow, and the pH value is 6.8, expand and cultivate continuously 3 times by volume ratio 1:20, obtain 1000ml of culture, Centrifuge at 12000rpm for 30min, and harvest 950ml supernatant for later use;

[0024] 2 Extraction of mycoplasma polysaccharide antigen

[0025] 2.1 Adj...

Embodiment 2

[0039]1 culture mycoplasma

[0040] 1.1 Culture medium preparation

[0041] Prepare 1500ml of modified KM2 medium. Mix 525ml of 1.7% hydrolyzed milk protein Hank's buffer solution, 30ml of 25% yeast infusion, 15ml of 1% thallium acetate solution, and 15ml of 0.4% phenol red. Membrane filtration sterilized minimum element medium (MEM) 750ml, healthy horse serum 150ml, 20,000 IU penicillin 15ml, adjusted the pH value to 7.8 with 1mol / L sodium hydroxide solution, and stored at 2°C for later use.

[0042] 1.2 Mycoplasma culture

[0043] Inoculate 5ml of the improved KM2 culture medium with Mycoplasma goatum subsp. goat pneumoniae, and culture it at 37°C for 5 days. 1200ml of the product was centrifuged at 12000rpm for 30min, and 1180ml of the supernatant was harvested for later use;

[0044] 2 Extraction of mycoplasma polysaccharide antigen

[0045] 2.1 Adjust the pH value of the culture supernatant to 5.0 with glacial acetic acid, boil for 60 minutes, filter with filter pape...

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Abstract

A mycoplasma capricolum subsp. pneumonia polysaccharide antigen of goats is characterized by extracting polysaccharides secreted by thalli from the supernatant of a mycoplasma capricolum subsp. pneumonia culture of goats. The agent is prepared by the following processing steps: a. culturing mycoplasma and b. extracting mycoplasma polysaccharide antigen. The invention has the following advantages:(1) the antigen in the invention is the polysaccharide antigen extracted from the supernatant of the mycoplasma capricolum subsp. pneumonia culture of goats and is polysaccharide secreted by thalli, has strong specificity and can truly detect the mycoplasma capricolum subsp. pneumonia antibody of goats; (2) the aldehyde-tanned sheep red cells sensitized with polysaccharide are used, thus avoidingthe defects of short preservation time and low sensitization titer of the fresh sheep red cells, being easily preserved for long time and having high hemagglutination titer;(3) the method is simple, convenient and fast in operation and dispenses with special equipment and apparatuses, thus being suitable for popularization and application at the grassroots level; (4) the invention is the unique diagnostic reagent for detecting the mycoplasma capricolum pneumonia antibodies of goats at home at present.

Description

technical field [0001] The present invention relates to the technical field of biological detection, in particular to a preparation method of Mycoplasma capricosum subsp. The invention discloses a method of indirect hemagglutination diagnostic reagent for detecting mycoplasma capricum subspecies capricum pneumoniae antibody. Background technique [0002] Mycoplasma capricosum goat pneumonia subspecies is the pathogen of goat infectious pleuropneumonia, which is one of the important diseases listed by the World Organization for Animal Health (OIE) and needs to be reported and monitored. Countries and regions that raise sheep bring serious economic losses to the sheep raising industry in these regions every year. The earliest written record of this disease in our country can be traced back to 1922 in the Yili area of ​​Xinjiang. Since then, more than ten provinces and cities including Gansu, Ningxia, Qinghai, Sichuan, Guizhou, Guangxi, Hebei, Liaoning, Inner Mongolia, Jiangsu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/04G01N33/531G01N33/86G01N33/569
Inventor 逯忠新储岳峰赵萍高鹏程贺英郭晗
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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