江西地区鼠源致病性钩端螺旋体菌的分离纯化与鉴定

Isolation,Purification and Identification of Pathogenic Leptospirosis from Rodents in Jiangxi Province

目的对江西地区钩端螺旋体菌进行分离纯化与鉴定,为后期建立江西地区钩端螺旋体菌的发病模型提供菌源。方法通过捕捉钩端螺旋体疫源地的储存宿主——褐家鼠,无菌操作摘取肾脏,将肾脏接种到EMJH培养基中培养、纯化菌株。对纯化后的钩端螺旋体用钩体外膜脂蛋白LipL32基因引物进行PCR检测以鉴定菌株,并对分离菌对金黄地鼠的致病性进行了研究。结果分离纯化到一株钩端螺旋体菌,PCR扩增为阳性,经核苷酸序列测定分析证实上述序列与钩端螺旋体菌FDAARGOS203基因序列(Gen Bank登录号:CP020414.2)同源性为100%。分离菌对金黄地鼠具有一定的致病性。结论我们成功从江西地区储存宿主褐家鼠肾脏中分离到一株具有致病性的钩端螺旋体菌株。本次研究为以后分离纯化钩端螺旋体菌累积了宝贵的经验,通过收集江西省内流行的钩端螺旋体菌型,为钩端螺旋体菌的发病模型提供菌源,并以此为基础,研发出可靠的临床早期诊断产品,以期达到有效控制江西省地区钩端螺旋体病的目的,降低公共卫生隐患。

Objective Separation,purification and identification of Leptospira in Jiangxi Province will provide a source of bacteria for the later establishment of the pathogenesis model of Leptospira in Jiangxi.Method The strain was obtained by aseptically removing the kidney from the storage host of the Leptospira source,the kidney,and inoculating the kidney into the EMJH medium to culture and purify the strain.The purified Leptospira was tested by PCR with the outer membrane lipoprotein LipL32 gene primer to identify the strain,and the pathogenicity of the isolate to golden hamster was studied.Result A strain of Leptospira was isolated and purified by PCR amplification.The nucleotide sequence analysis confirmed that the sequence was 100%homologous to the Leptospira FDAARGOS203 gene sequence(GenBank accession number:CP020414.2).The isolate has certain pathogenicity to the golden hamster.Conclusion We successfully isolated a pathogenic strain of Leptospira from the kidney of the storage rat brown rat.This study has accumulated valuable experience for the separation and purification of Leptospira in the future.By collecting the popular Leptospira type in Jiangxi Province,it provides a source of bacteria for the pathogenesis model of Leptospira,and based on this,developed reliable clinical early diagnosis products,in order to achieve effective control of leptospirosis in Jiangxi Province,and reduce public health hazards.