首页> 中文期刊> 《中国人兽共患病学报》 >台湾海峡异尖线虫病病原线虫幼虫的rDNA序列分析

台湾海峡异尖线虫病病原线虫幼虫的rDNA序列分析

         

摘要

To analyze the rDNA sequence of the Anisakid nematodes with zoonotic potential from Taiwan straits, Anisakid nematodes were obtained from the gut of marine fish and identified chiefly based on the morphological characteristics. The genomic DNA of nematodes were extracted, and the sequence of rDNA ITS were amplified by PCR using universal primers, then cloned and sequenced bidirectionally. Sequences analysis was conducted by blasting database and software DNAMAN. Results of the blasting database showed that no match could be demonstrated in 3 of 6 rDNA sequences obtained from 6 different nematode species;the new sequences were submitted to the database and the accession numbers in GenBank were obtained. The relationship between adult Contracaecum muraenesoxi and its putative third stage larva was validated by molecular evidences. Sequences relationship based on the inner similarity of the internal transcribed spacer sequences was constructed. It was found that the phylogenetic informative loci were mainly gathered in the second internal transcribed spacer(ITS2) for these anisakid nematodes because of the ITS2 has higher transition ratio than that of the ITS1. It is evident that the sequence in this region could provide valuable information for the molecular distinction of anisakid nematodes with zoonotic potential from Taiwan straits.%目的 分析台湾海峡异尖线虫病潜在病原线虫的rDNA序列,为建立病原分子生物学鉴别提供依据.方法 剖检台湾海峡常见鱼类的寄生线虫幼虫,镜检分类.提取虫体DNA、用通用引物对rDNA进行PCR扩增、克隆、双向测序及BLAST比对.结果 获得6条来自6种线虫幼虫的rDNA序列,其中的3种在GenBank数据库中没有发现匹配的序列;证实在台湾海峡分布的灰海鳗对盲囊线虫成虫和疑似其第三期幼虫实属同种;构建了基于ITS-2的序列间内部相似性程度关系树.结论 异尖线虫病原幼虫所具有的分子生物学鉴别信息位点主要集中在ITS-2间隔区.

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