鉴别MS-H疫苗株和MS野毒株的MAMA-PCR检测方法的建立与初步应用

Establishment and preliminary application of MAMA-PCR method for identification of MS-H vaccine strain and MS wild strain

根据Kreizinger Z等[1]已发表的研究,依据obg基因第367位和629位的核苷酸转换,我们建立了依托琼脂糖凝胶电泳(常规PCR)的错配扩增突变分析(MAMA)方法以区分ts+MS-H疫苗株、ts-MS-H再分离株以及MS野毒株。本研究对本实验室保存的客户送检的MS分离株的阳性病料,以及日常场区送检咽喉拭子MS通用引物检测的阳性样品进行MAMA-PCR扩增,用以比对各分离株目的片段的差别,建立MS-H疫苗株与MS野毒株的鉴别方法,这将为临床鸡群MS感染状态的评估提供参考依据,也为MS的净化和防控提供技术指导。

According to the published studies of Kreizinger Z[1],based on the substitutions at nt367 and nt629 of the obg gene,we established a mismatch amplification mutation analysis(MAMA)method based on agarose gel electrophoresis(conventional PCR)to distinguish the ts+MS-H vaccine strain genotype from its non-temperature sensitive re-isolate genotype and wild-type M.synoviae isolate genotype.In this study,MAMA-PCR was used to amplify the positive samples of MS isolates stored in our laboratory and those of throat swabs tested by MS universal primers in daily field.To compare the differences of the target fragments of each isolate,the identification method of MS-H vaccine strain and MS wild strain was established to provide a reference for the evaluation of MS infection status in clinical chicken flocks and technical guidance for the purification and control of MS.