胞内劳森菌巢式PCR检测方法的建立及初步临床应用  

Development and preliminary application of a nested PCR menthod for detection of Lawsonia intracellularis in feces of pigs

作　　者：彭忠[1] 涂志勤[1] 梁婉[1] 胡睿铭[1] 王豪男 寿琎 董超[1] 李宗华[1] 陈焕春[1] 吴斌[1] 

机构地区：[1]华中农业大学动物医学院/农业微生物学国家重点实验室/生猪健康养殖协同创新中心,湖北武汉430070

出　　处：《中国兽医学报》2017年第1期40-46,共7页Chinese Journal of Veterinary Science

基　　金：国家科技支撑计划资助项目(2014BAD20B00);湖北省科技支撑计划资助项目(2013BBB14)

摘　　要：为了掌握胞内劳森菌在湖北省5个疑似猪增生性肠炎暴发规模化猪场中的流行情况,我们建立了一种从粪便中检测胞内劳森菌的巢式PCR方法,并利用该方法对采集于这5个规模化猪场不同生产阶段不同临床表现猪群的1 003份粪便样品进行监测。结果显示:5个猪场的胞内劳森菌检出率在11.5%~19.4%之间,平均阳性率为15.5%;就临床表现而言,腹泻猪粪便样品中胞内劳森菌的检出率高于非腹泻猪;就生产阶段而言,保育及育肥猪中胞内劳森菌的检出率高于断奶仔猪、母猪和公猪。我们还利用粪便检测胞内劳森菌呈阳性、疑似临床症状明显的猪的肠道制备的感染溶液进行了病例复制,试验猪在灌服感染溶液后出现了与胞内劳森菌感染相似的临床症状及病理变化,并且从感染猪的肠道组织和粪便中检测到了胞内劳森菌的存在。本试验对于进一步了解胞内劳森菌在我国猪群中的流行情况及致病性具有一定的意义。To investigate the prevalence of Lawsonia intracellularis in five intensive pig farms in which a porcine proliferative enteropathy （PPE） like disease appeared at the same period in 2014, we developed a nested PCR menthod to detect L. intracellularis in feces of pigs. A total of 1 003 fecal samples from pigs of different ages in these farms were collected and detected. Results re- vealed 15.5%（155/1 003） average positivity in the five farms and the positive rate varied from 11.5% （23/200） to 19.4% （35/180）. The positive rates were higher in grower-finisher pigs （6-20 weeks old） than that of other ages,and a similar result appeared in diarrheic pigs compared with the normal pigs. Besides, we reproduced the infection of L. intracellularis in healthy piglets via administered orally with a total of 60 mL affected mucosa homogenates. PPE similar clinical signs and pathological changes occurred after infection, and the nucleotide of L. intracellularis was de- tected from the ileal and fecal samples of the challenged pigs. Our work would have a contribution to understanding the prevalent condition and the pathogenesis of L. intracellularis.

关 键 词：胞内劳森菌 猪群 阳性率 巢式PCR 病例复制 

分 类 号：S852.61[农业科学—基础兽医学]