3种禽支原体多重PCR检测方法的建立及应用

Establishment and Application of Multiple PCR for Diagnosing Three Mycoplasmas

从鸡毒支原体、鸡滑液囊支原体和火鸡支原体3种致病性支原体液体培养物中提取DNA,用特定引物分别和组合进行PCR,均得到了特异性扩增产物,片段大小分别为732、207和850bp。直接用液体培养物进行PCR亦得到了一致的电泳条带。试验结果表明,建立的PCR方法可用于上述3种支原体临床感染的鉴别诊断。ELISA血清学检测和气管拭子PCR检测的比较结果表明,该PCR方法可用于临床MG检测。

DNA of three types of avian Mycoplasma（Mycoplasma gallisepticum,MG;Mycoplasma synoviae,MS and Mycoplasma meleagridis,MM） were extracted and amplified through single polymerase chain reaction（PCR） and multiplex PCR with three sets of specific oligonucleotide primers.There were consistent specific electrophoresis bands by both single PCR assay and multiplex PCR assay,and the fragments were gained respectively,that was 732 bp for MG,207 bp for MS and 850 bp for MM.While the same bands were gained on agarose gel,the fluid cultures containing Mycoplasma were directly performed for PCR.These data showed that the established multiplex PCR assay might facilitate differentiation of clinical infection by different serotype of avian Mycoplasma.At the same time,the comparison test between ELISA for serum and PCR for MG in chickens also presented the availability of detecting MG clinically.