云贵两省三地带绦虫的分子鉴定——核糖体DNA第一内转录间隔区(ITS1)限制性酶切片段长度多态性(RFLP)分析

MOLECULAR IDENTIFICATION OF TAENIA IN THREE REGIONS OF YUNNAN AND GUIZHOU PROVINCES——RESTRICTION FRAGMENT LENGTH POLYMORPHISM(RFLP) ANALYSIS OF THE FIRST INTERNAL TRANSCRIBED SPACER(ITS1) OF RIBOSOMAL DNA

目的用PCR-RFLP方法对rDNA-ITS1片段进行分析,以进一步明确云贵地区是否存在牛带绦虫亚洲亚种,并建立一种快速鉴定方法。方法取贵州都匀株(DY)、贵州从江株(CJ)、云南大理株(DL)带绦虫及台湾株(TW)成虫标本,剪取孕节,抽提DNA,PCR扩增rDNA-ITS1片段,分别用4种限制性内切酶MspI、CfoI、AluI、RsaI对扩增片段作酶切分析。结果PCR产物经AluI、RsaI酶切后,TW、DL、DY和CJ株RFLP图谱一致;经MspI、CfoI酶切后,TW、DL和DY株RFLP图谱一致,CJ株显著不同。结论1)DL和DY株与TW株同属牛带绦虫亚洲亚种;而CJ株是传统牛带绦虫;2)rDNA-ITS1的PCR-RFLP分析方法简便,可以用于带绦虫的分类学研究。

Objective To further tell whether there was Taenia saginata asiatica in Guizhou and Yunnan Province with PCRRFLP of ITS1 and to establish a simple method to distinguish both Taenia. Methods Genomic DNA was extracted from the adult samples of Yunnan Dali（DL）, Guizhou Duyun（DY）, Guizhou Congjiang（CJ） and Taiwan（TW） ; rDNA-ITS1 was amplified with universal primers and the amplified fragments were digested with the 4 restriction endonucleases （MspI.CfoI.AluI.RsaI） respectively for analysis. Results Identical electrophoresis maps were found in the TW, DL, CJ and DY strains when with AluI and RsaI; and it was distinct restriction patterns between the CJ strain and the other strains （DY, DL, TW） when with MspI and CfoI. Conclusion 1） DL, DY and TW strains belong to Taenia saginata asiatica and CJ strain is typical Taenia saginata ; 2） PCR-RFLP of rDNA-ITS1 is a simple method for taxological study of Taenia.