PCR检测感染早期小鼠血液中旋毛虫幼虫的研究  

Early detection of Trichinella spiralis larvae by the polymerase chain reaction in blood of the infected mice

作　　者：李峰[1] 王中全[1] 崔晶[1] 王书伟[1] 李楠[1] 

机构地区：[1]郑州大学医学院寄生虫学教研室,河南郑州450052

出　　处：《中国预防兽医学报》2009年第12期936-940,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：河南省重大公益性科研项目(2008);河南省重点科技攻关项目(092102110091);河南省医学科技攻关项目200803001)

摘　　要：为了研究PCR检测感染小鼠血液中旋毛虫DNA的敏感性,应用旋毛虫1.6kb重复序列为扩增靶序列对旋毛虫(T1)、乡土旋毛虫(T2)、布氏旋毛虫(T3)、伪旋毛虫(T4)和南方旋毛虫(T7)肌幼虫DNA进行PCR扩增,并检测小鼠感染20、100、300条T1肌幼虫后不同时间的外周血。结果表明,T1、T4和T7肌幼虫可扩增出特异性目的条带(510bp),而T2和T3无扩增产物;1、0.04和0.02条T1、T4和T7肌幼虫均能扩增到清晰的目的条带(510bp)。20条幼虫感染小鼠后5d～6d,PCR阳性率均为7.69%;100条幼虫感染小鼠后5d～12d可检出旋毛虫DNA,其中感染后5d～7d的阳性率分别为30.77%、38.46%及30.77%;300条幼虫感染小鼠后5d～15d可检出旋毛虫DNA,感染后7d的阳性率为61.54%,感染后6d与8d～10d的阳性率均为53.85%。3组旋毛虫感染小鼠PCR阳性率间的差异有统计学意义(p<0.01),PCR阳性率随感染剂量的增加而升高(p<0.01),100条与300条感染小鼠感染后不同时间的PCR阳性率与检测时间有相关性(p<0.01)。以上实验结果表明PCR检测感染小鼠血液中旋毛虫DNA的敏感性与感染程度和检测时间有关,对感染早期旋毛虫抗体阴性宿主有一定诊断价值。To detect migratory larvae at early stage of Trichinella spiralis infection, PCR was developed to detect muscle larvae DNA of T. spiralis, T. nativa, T. britovi, T. pseudospiralis and T. nelson using primers for amplification of 1.6 kb repetitive sequence of T. spiralis. A PCR fragrnent of 510 bp was amplified from DNA of T. spiralis, T. pseudospiralis and T. nelsoni, but no products amplified from T. nativa and T. britovi. Mice were infected with 20, 100 or 300 T. spiralis larvae and Trichinella-specific DNA in peripheral blood was detected by PCR at different days post infection （dpi）. T. spiralis DNA could be detected in the infected mice from 5 dpi, with a detection rate from 7.69 % in mice infected with 20 larvae to 61.54 % in mice infected with 300 larvae. The PCR positive rate elevated with the increase of infecting dose （p〈0.01）. In mice infected with 100 and 300 larvae, PCR positive rates showed significant positive correlation with the detection time after infection （p 〈 0.01）. The sensitivity of PCR detecting DNA of T. spiralis migratory larvae in blood of infected mice depended on the severity and stage of infection.

关 键 词：旋毛虫 移行期幼虫 PCR 

分 类 号：S852.71[农业科学—基础兽医学]