Preliminary Application And Estanlish Of PCR Assay For Detection Of Clostridium Chauvoei

Emphysematous gangrene caused by clostridium emphysematous gangrene was a hot given priority to with cattle, acute, local infection disease. Pathogens for clostridium emphysematous gangrene. Main symptom was that the color of the dry skin of muscle disease dark red or black. Cut the skin, outflowing black red liquid with sour smell flowing from the incision. Mainly through the mouth and throat trauma intrusion into organization, also could be invaded blood by flabby gastric mucosa or the micro injury. Then given rise to systemic infection. Due to the speed of clostridium emphysematous gangrene growth relatively slow compared with other clostridium, because of its easily happened cross infection with other clostridium, so it was difficult to accurately detection by the perspective of the etiology and immunology. Cattle Emphysematous gangrene was easily to form spore. It was insensitive to acid, alkali, temperature as well as universal disinfectant as not sensitive, Emphysematous gangrene could survive for 5 years in the soil especially, indeed survive for 10 years or longer in dry epidemic materials. Emphysematous gangrene clostridium usually harmed cattle,It was more susceptible to the fungus than other: animals, and developed in calves under the age of three, sheep and buffalo also infected emphysematous gangrene.It had great harm to livestock and human. So it was particularly important to establish a fast and efficient detection method. In recent years, with the continuous development of biological research and technology, and gradually established a new serological methods of molecular biology diagnosis. In the short term, the detection method of emphysematous gangrene mainly included the separation identification as well as clostridium polymerase chain reaction (PCR),.Separation identification was a method which detection rate was extremely low and complicately. Testing by PCR technology, it was sensitive, rapid, and had high specificity, PCR is a valuable for the emphysematous gangrene diagnosis and research. Polymerase chain reaction (PCR),by the American Kary Mullis, etc have pioneered and developed by Cetus companies in the United States. It was a laboratory method for amplification DNA fragments In vitro. Up to now, PCR technology made great progress and constantly improved the reliability, many technologies regarded PCR technology as the foundation had been quite objective development.The experiment for rapid detection methods of Clostridium emphysematous gangrene (Clostridium chauvoei), according to the GenBank gas gangrene swoll en Clostridium cytotoxic cctA gene sequences (registration number:JQ692583) designed a pair of primers, Clostridium emphysematous gangrene standard strai ns C54-1 whole genome DNA as a template, set up a cattle emphysematous g angrene PCR detection method, provided basic research for rapid detection an d diagnosis of cattle emphysematous gangrene.The results showed that established PCR amplification fragment size of emp hysematous gangrene was 501 bp, and clostridium emphysematous gangrene on GenBank homology was 99.4%,there was no cross reaction with type D,type E, clostridium corruption and pasteurella, the minimum detectable concentration is 12.30 fg/mu L. The results showed that the established PCR detection meth od hasmore suitable for the diagnosis of clostridium emphysematous gangrene. The advantage was sensitive and efficient compared with other diagnosis metho d.