云南6地域带绦虫mtCOXⅠ片段序列测定分析

目的利用mtCOXⅠ片段的PCR和序列测定鉴定云南6地域（怒江、大理、迪庆、西双版纳、临沧、楚雄）带绦虫的虫种,并构建系统发育树。方法从6地域采集带绦虫成虫,抽提DNA,PCR扩增mtCOXⅠ片段,并做序列测定;结合GenBank中已知的猪带绦虫、牛带绦虫、亚洲带绦虫mtCOXⅠ序列,经DNA MAN软件处理后计算遗传距离并构建系统发育树状图。结果遗传距离矩阵表显示：NJ4、NJ1和DQ2的遗传距离为99.8%,DL4和NJ3的遗传距离为99.5%,NJ2和DQ3的遗传距离为98.8%,DL3与BZ3的遗传距离为98.3%,而与BZ2的遗传距离为96.0%;系统发育树显示：NJ1～4、DL2～3及DQ1～3共10个标本株与BZ3聚为一支.BN1、CX1、LC1与BZ2聚为一支,两支交汇后再与由DL1和BZ1聚合的一支汇合。结论怒江和迪庆各株为亚洲带绦虫,西双版纳、临沧和楚雄株为牛带绦虫,大理株为亚洲带绦虫和猪带绦虫。同虫种不存在地理区域的差异。mtCOXⅠ片段的序列测定可用于带绦虫分类鉴定。

Sequence analysis on mt COXI fragment of Taenia cestodes from 6 regions of Yunnan province

To identify Taenia cestodes from 6 regions of Yunnan province by PCR and sequencing of mtCOXⅠ fragment. the genomic DNA of Taenia cestodes was extracted from proglottid collected in 6 region of Yunnan province, and mtCOXⅠ gene fragments were amplified by PCR, and then sequenced. The genomic distance and phylogenetic tree were constructed in comparison with other known mtCOXⅠ gene sequences of T. solium , T. saginata and T. asiatica in GenBank using DNA MAN software. Through distance matrix,it was found that the homologie of NJ4, NJ1 and DQ2 was 99.8 %, DL4 and NJ3 homoiogie was 99.5%, NJ2 and DQ3 homologie was 98.8%; the homologie of DL3 and BZ3 was 98.3%, while the homologie was 96.0 % with BZ2; The phylogenetic tree demonstrated that 10 Taenia cestodes including NJ1-4, DL2-3and DQ1-3 occupied one brance with BZ3. BN1, CX1, LC1 and BZ2 occupied one brance, then two brance occupied and occupied with other one which was occupied by DL1 and BZ1. Taenia cestodes from Nujiang and Diqing were T. asiatica. Taenia cestodes from XiShangbanna, Lincang and Chuxiong were T. saginata. Taenia cestodes from Dali were T. solium or T. asiatica. Because same species have no difference from different regions, mtCOXⅠ fragment sequencing is valid for tapeworms identification.