【摘要】 目的对我国旋毛虫不同地理株进行分子鉴定。方法根据旋毛虫线粒体细胞色素氧化酶Ⅰ(cytochromec-oxidase subunitⅠ,COXⅠ)基因序列合成1对引物,以旋毛虫(Trichinella spiralis,T1)、乡土旋毛虫(T.nativa,T2)、布氏旋毛虫(T.britovi,T3)、伪旋毛虫(T.pseudospiralis,T4)、纳氏旋毛虫(T.nelsoni,T7)的国际参考株作为对照,应用PCR-限制性片段长度多态性(PCR-restrictionfragment length polymorphism,PCR-RFLP)技术对我国7个猪源旋毛虫地理株(河南、湖北、云南、西安、哈尔滨、同江及天津株)进行分子鉴定。结果T1、T2、T3、T4、T7和我国7个猪源旋毛虫地理株的COXⅠ基因均扩增出419 bp的片段,PCR扩增产物Tru1Ⅰ(MseⅠ)酶切后经RFLP分析,T2、T3、T4、T7和天津株均具有独特的酶切带型:T2为22、70和327 bp,T3为92和327 bp,T4为419 bp,T7为62、64、70和223 bp,天津株为92、126和201 bp;其他6个地理株的酶切带型和T1一致,为22、70、126和201 bp。结论我国7个旋毛虫地理株的COXⅠ基因片段(92、70和22 bp)存在种内多态性;河南、湖北、云南、西安、哈尔滨及同江株可归为一类,天津株可归为另一类。更多还原

【Abstract】 Objective To identify the different Trichinella isolates from China at molecular level.Methods One pairs of primers were synthesized based on the DNA sequence of mitochondrial cytochrome c-oxidase subunitⅠ(COXⅠ) gene of Trichinella.Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) was applied for the molecular identification of seven swine Trichimella isolates from Henan,Hubei,Yunnan,Xi’an,Harbin,Tongjiang and Tianjin.International reference strains of five species of Trichinella [T.spiralis(T1),T.nativa(T2),T.britovi(T3),T.pseudospiralis(T4)and T.nelsoni(T7)]were used as control.Results The results of electrophoresis of PCR products of Trichinella COXⅠ genes showed that the band(419 bp) of international reference strains of five species of Trichinella(T1,T2,T3,T4 and T7) was the same as seven Trichinella isolates from China.Electrophoresis of COXⅠ gene PCR products digested by restriction endonuclease Tru1Ⅰ(MseⅠ) showed that T2,T3,T4,T7 and Tianjin isolate have their restriction fragments: T2(22 bp,70 bp,327 bp),T3(92 bp,327 bp),T4(419 bp),T7(62 bp,64 bp,70 bp,223 bp) and Tianjin isolate(92 bp,126 bp,201 bp).The other six isolates and T1 have the same restriction fragments(22 bp,70 bp,126 bp,201 bp).Conclusion This study reveals an intraspecies polymorphism in the 92 bp,70 bp and 22 bp fragments of COXⅠ gene of seven T.spiralis isolates from China.The isolates from Henan,Hubei,Yunnan,Xi’an,Harbin,Tongjiang are classified as one group,the Tianjin isolate is classified as another group.更多还原