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尼日利亚西南部鉴定泡螺属物种的分子方法及对血吸虫自然感染蜗牛的观察

Molecular approaches to the identification of Bulinus species in south-west Nigeria and observations on natural snail infections with schistosomes.

作者信息

Akinwale O P, Kane R A, Rollinson D, Stothard J R, Ajayi M B, Akande D O, Ogungbemi M O, Duker C, Gyang P V, Adeleke M A

机构信息

Molecular Parasitology Laboratory, Public Health Division, Nigerian Institute of Medical Research, Yaba, Lagos, Nigeria.

出版信息

J Helminthol. 2011 Sep;85(3):283-93. doi: 10.1017/S0022149X10000568. Epub 2010 Sep 21.

Abstract

The current study considers the distribution of a small sample of 138 Bulinus snails, across 28 localities within eight Nigerian states. Snails were identified using a combination of molecular methods involving both DNA sequencing of a partial cytochrome oxidase subunit 1 (cox1) fragment and restriction profiles obtained from ribosomal internal transcribed spacer (its) amplicons. The results showed that the majority of Bulinus samples tested belonged to the species Bulinus truncatus while only two were Bulinus globosus. The use of RsaI restriction endonuclease to cleave the ribosomal its of Bulinus, as a method of species identification, was adopted for the majority of samples, this being a quicker and cheaper method better suited to small laboratory environments. Polymerase chain reaction (PCR) amplification of the schistosome Dra1 repeat within each of the collected Bulinus samples was employed to determine the extent and distribution of infected snails within the sample areas. Successful amplification of the Dra1 repeat demonstrated that 29.7% of snails were infected with schistosomes. Sequencing of the partial schistosome its from a small subset of snail samples suggested that some snails were either penetrated by both Schistosoma haematobium and Schistosoma bovis miracidia or hybrid miracidia formed from the two species.

摘要

本研究考察了138只布氏螺的小样本在尼日利亚八个州的28个地点的分布情况。通过分子方法组合来鉴定螺类,这些方法包括对细胞色素氧化酶亚基1(cox1)部分片段进行DNA测序以及从核糖体内部转录间隔区(its)扩增子获得限制性图谱。结果显示,所检测的大多数布氏螺样本属于截形布氏螺物种,而只有两个是球茎布氏螺。大多数样本采用RsaI限制性内切酶切割布氏螺核糖体its作为物种鉴定方法,这是一种更快速、更便宜的方法,更适合小型实验室环境。对每个采集的布氏螺样本中的血吸虫Dra1重复序列进行聚合酶链反应(PCR)扩增,以确定样本区域内受感染螺的范围和分布。Dra1重复序列成功扩增表明29.7%的螺感染了血吸虫。对一小部分螺样本的部分血吸虫its进行测序表明,一些螺要么被埃及血吸虫和牛血吸虫的毛蚴同时穿透,要么被这两个物种形成的杂交毛蚴穿透。

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