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利用DNA探针进行乌干达盘尾丝虫的虫株鉴别

Strain differentiation of Onchocerca volvulus from Uganda using DNA probes.

作者信息

Fischer P, Bamuhiiga J, Kilian A H, Büttner D W

机构信息

Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.

出版信息

Parasitology. 1996 Apr;112 ( Pt 4):401-408. doi: 10.1017/s0031182000066634.

Abstract

Polymerase chain reaction (PCR) combined with non-radioactive DNA hybridization was applied for the detection and characterization of a 150 bp tandem repeat of Onchocerca volvulus. DNA of worms from western Uganda was amplified and then probed with a digoxygenin-labelled oligonucleotide, specific for the forest form of O. volvulus and compared to samples from various African countries. Hybridization was only observed with PCR products from the forest in Liberia, south-eastern Ghana, Benin and southern Cameroon, but not with worms from Uganda or the savannah in Burkina Faso and northern Ghana. A nested PCR using primers derived form the forest form-specific DNA sequence confirmed these results. Morphometric studies revealed length differences between the microfilariae of Ugandan O. volvulus to those of West Africa, especially to those of the savannah in Burkina Faso. It is concluded that the forest/savannah classification of O. volvulus from West Africa is not suitable for Simulium neavei-transmitted O. volvulus from Uganda.

摘要

聚合酶链反应(PCR)结合非放射性DNA杂交技术被用于盘尾丝虫150碱基对串联重复序列的检测与特征分析。对来自乌干达西部的虫体DNA进行扩增,然后用针对盘尾丝虫森林型的地高辛标记寡核苷酸进行探针杂交,并与来自非洲各国的样本进行比较。仅在来自利比里亚森林地区、加纳东南部、贝宁和喀麦隆南部的PCR产物中观察到杂交信号,而来自乌干达的虫体或布基纳法索和加纳北部草原地区的虫体则未出现杂交信号。使用源自森林型特异性DNA序列的引物进行巢式PCR证实了这些结果。形态测量研究表明,乌干达盘尾丝虫的微丝蚴与西非的微丝蚴在长度上存在差异,尤其是与布基纳法索草原地区的微丝蚴。得出的结论是,西非盘尾丝虫的森林型/草原型分类不适用于乌干达由纳氏蚋传播的盘尾丝虫。

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