摘要
为快速检测牛支原体(M.bovis),根据GenBank中登录的M.bovis P81基因序列设计特异性引物,通过条件优化,建立了M.bovis环介导等温扩增(LAMP)检测方法。将该方法的灵敏度与巢式PCR进行对比,并用其检测牛支原体外的其他几种病原菌,同时对疑似感染M.bovis的临床病料进行检测。该方法的灵敏度比巢式PCR高103倍,最低检出限为1.1ng/L,对其他几种病原菌的检测结果均为阴性,临床病料检测结果与分离培养符合率为100%。建立的M.bovis LAMP检测方法灵敏度高、特异性强、检测快速准确,且不需要专门的仪器设备,可用于牛支原体病的临床检测。
In order to detect M. boris rapidly, the specific primers were designed according to the GenBank registered M. boris P81 gene sequence to establish M. boris loop mediated isothermal am- plification （LAMP） detection method by optimizing condition. The sensitivity of this method were compared with nested PCR,several other pathogens and clinical samples suspected infection M. bo- ris were detected by this method. The sensitivity of this method was 103 times higher than the nes- ted PCR,its detection limit was 1.1 ng/L. The test results were negative for several other patho- gens,and the coincidence rate of clinical samples test results and isolation was 100%. The M. boris LAMP detection method is rapid and accurate and has high sensitivity and specificity,and does not require specialized equipment, so it can be used for the clinical detection of M. boris disease.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2014年第4期571-577,582,共8页
Chinese Journal of Veterinary Science
基金
国家科技支撑计划课题资助项目(2012B-AD43B02)
石河子市农业科技攻关计划资助项目(2011NY10)
