Otranto D, Testini G, De Luca F, Hu M, Shamsi S, Gasser R B
Department of Animal Health and Welfare, University of Bari, Faculty of Veterinary Medicine, P.O. Box 7, Str. Prov. per Casamassima Km3, 70010 Valenzano, Bari, Italy.
Mol Cell Probes. 2005 Oct;19(5):306-13. doi: 10.1016/j.mcp.2005.05.001.
This study investigated genetic variability within the 'eyeworm'Thelazia callipaeda (Nematoda: Thelazioidea) from Europe and Asia by polymerase chain reaction (PCR)-coupled sequencing and mutation scanning of the mitochondrial cytochrome c oxidase subunit 1 gene (cox 1). Eight different sequence variants of cox 1 (haplotypes) were determined for the 50 individual adult specimens of T. callipaeda (from dogs, foxes or cats from Italy, Germany and the Netherlands and from dogs from China and Korea). Nucleotide variation (0.3--2%) was detected at 23 of 649 positions in the cox 1. Six of these positions were invariable among all 37 individuals from Europe and among the 13 individuals from Asia (irrespective of host origin) but differed (five G<-->A and one C<-->T changes) between Europe and Asia. PCR-based single-strand conformation polymorphism (SSCP) analysis of the most variable portion (v-cox 1) of the cox 1 was validated (for a subset of samples) as a tool to rapidly screen for genetic (haplotypic) variability. The results for the SSCP analysis and sequencing were concordant, indicating that the mutation scanning approach provides a useful tool for investigating the population genetics and molecular ecology of T. callipaeda.
本研究通过聚合酶链反应（PCR）结合线粒体细胞色素c氧化酶亚基1基因（cox 1）的测序及突变扫描，调查了来自欧洲和亚洲的“眼线虫”结膜吸吮线虫（线虫纲：吸吮总科）的遗传变异性。对50个结膜吸吮线虫成虫样本（来自意大利、德国和荷兰的犬、狐或猫，以及来自中国和韩国的犬）测定了cox 1的8种不同序列变体（单倍型）。在cox 1的649个位置中的23个位置检测到核苷酸变异（0.3% - 2%）。其中6个位置在来自欧洲的所有37个个体以及来自亚洲的13个个体（不论宿主来源）中是不变的，但在欧洲和亚洲之间存在差异（5个G<-->A和1个C<-->T变化）。基于PCR的单链构象多态性（SSCP）分析cox 1最可变部分（v - cox 1）（针对部分样本）被验证为快速筛选遗传（单倍型）变异性的工具。SSCP分析和测序结果一致，表明突变扫描方法为研究结膜吸吮线虫的群体遗传学和分子生态学提供了有用的工具。
