猪肺炎支原体PCR快速检测方法的应用研究

Application research of rapid detection method in Mycoplasma hyopneumoniae

为对猪场中猪肺炎支原体的感染和发病情况进行有效地病原学监测,本研究筛选国内外最常用且引用率最高的靶向16S rRNA基因的引物对作为检测猪肺炎支原体的特异性引物,通过制定临床猪肺组织样品、鼻拭子样品和肺泡灌洗液样品的标准化采集与处理方法,比较菌液样品、肺组织样品、鼻拭子样品与支气管肺泡灌洗液等不同样品的不同DNA提取方法,验证PCR检测方法适用的临床样本范围。结果表明,对于菌液样品和鼻拭子样品可选用经济快速的水煮法,对于肺组织样品和肺泡灌洗液样品使用高效的试剂盒提取法。本研究表明,使用靶向16S rRNA基因的引物对进行PCR检测,可满足临床上对猪肺炎支原体快速检测的要求。

In order to investigate the potential involvement of gycoplasma hyopneumoniae in porcine respiratory disease in pigs efficiently,a primer set of 16S rRNA,which has been wildly used, was used to monitor the incidence of Mycoplasma hyopneumoniae infection in herds clinically. This paper set gathering and dealing methods for PCR applicable standardized clinical samples for pig lung tissues,nasal swabs and bronchoalveolar lavage fluids. Meanwhile,the study compared different DNA extracting methods for different samples,such as bacteria fluid sample,lung tissues,nasal swabs and bronchoalveolar lavage fluids,etc. It was acknowledged that one could use the economic and rapid method-boiling method for bacteria fluid samples and nasal swabs samples,while one could use efficient extraction kit method for lung tissues and bronchoalveolar lavage fluids. Clinical application has proved that this PCR method satisfies the clinical requirements of detecting Mycoplasma hyopneumoniae rapidly.