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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。
Objective To identify the different geographical strains of Trichinella in China. Methods A pair of primers was synthesized based on the sequence of mitochondrial cytochrome c-oxidase subunit Ⅰ (COX Ⅰ) in Trichinella spiralis (T), Trichinella spiralis (T1), T. nativa (T2) T. tuberculosis (T. tvitivi, T3), T. pseudospiralis (T4) and T.nelsoni (T7) were used as controls. The PCR-restriction fragment length polymorphism restriction fragment length polymorphism and restriction fragment length polymorphism (PCR-RFLP) were used to identify the seven Trichinella spiralis geographic isolates (Henan, Hubei, Yunnan, Xi’an, Harbin, Tongjiang and Tianjin). Results A 419 bp fragment was amplified from T1, T2, T3, T4, T7 and COX Ⅰ gene of 7 Trichinella spiralis geographic isolates. RFLP analysis and RFLP analysis showed that the product of PCR amplification was Tru1Ⅰ (MseⅠ) T4, T7 and Tianjin strains had unique digestion bands: T2 was 22, 70 and 327 bp, T3 was 92 and 327 bp, T4 was 419 bp, T7 was 62, 64, 70 and 223 bp, respectively 92, 126 and 201 bp, respectively. The other 6 geographical isolates were consistent with T1, 22, 70, 126 and 201 bp. Conclusion There are intraspecific polymorphisms in the COXⅠ gene fragments (92, 70 and 22 bp) in seven Trichinella strains in China. Henan, Hubei, Yunnan, Xi’an, Harbin and Tongjiang can be classified as one species, For another class.