Establishment and Application of Nested PCR Detection Method of Pathogens of Mycoplasma ovipneumonia
[Objective]In order to establish a nested polymerase chain reaction(PCR)for detecting the pathogens of Mycoplasma ovipneumo-nia,and determine the target organs of mycoplasma infection.[Method]According to the 16S rRNA sequence of sheep mycoplasma in Gen-Bank website,two pairs of specific primers were designed and synthesized.PCR reaction conditions were optimized by using the genome DNA of M.ovipneumonia as templates.The nested PCR products were verified by sequencing.A nested PCR method for detecting the pathogens of M.ovipneumonia was established.The established method was used to detect the lung,lung lymph,heart,kidney,liver,spleen,skin,small intestine and peripheral blood of clinical positive samples,and the lung tissue from clinical suspected samples.[Result]864 bp specific gene fragment was amplified by using nested PCR method.The lung and lung lymph of sheep were the target organs of mycoplasma infection.The coincidence rate between the detection rate of clinical samples by the nested PCR and pathogen culture and identification results was 100％.[Conclusion]The nested PCR method for detection of M.ovipneumonia could be applied for the laboratory diagnosis of clinical samples.
万方数据
维普
