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基于核糖体DNA内转录间隔区-1序列可变简单序列重复通过多重PCR鉴定瘿螨(蜱螨亚纲:瘿螨科)

Identification of cecidophyopsis mites (Acari: Eriophyidae) based on variable simple sequence repeats of ribosomal DNA internal transcribed spacer-1 sequences via multiplex PCR.

作者信息

Lava Kumar P, Fenton B, Jones A T

机构信息

International Crops Research Institute for the Semi-Arid Tropics, Patancheru, Hyderbad, India.

出版信息

Insect Mol Biol. 1999 Aug;8(3):347-57. doi: 10.1046/j.1365-2583.1999.83123.x.

Abstract

A PCR multiplex technique was developed for identifying Cecidophyopsis mites using species-specific differences in rDNA ITS-1 sequences. Four PCR primers derived from ITS-1 were used for the simultaneous amplification (multiplex PCR) of interspecifically variable simple sequence repeats (vSSRs). Mites were identified by electrophoresing PCR products alongside those obtained from plasmids containing ITS copies of known mite species. The multiplex PCR assay was rapid, reproducible and had a sensitivity comparable to sequencing. It was used to identify mite specimens on Ribes from around the world. It also identified a profile from mites on R. rubrum that had no equivalent amongst the known Cecidophyopsis species. Sequence and ecological analysis of this mite suggest that it is a new species of nongall-forming Cecidophyopsis mite.

摘要

开发了一种聚合酶链式反应(PCR)多重技术,用于利用核糖体DNA内转录间隔区1(rDNA ITS-1)序列的种间差异来鉴定瘿螨。从ITS-1衍生的四种PCR引物用于同时扩增种间可变简单序列重复序列(vSSRs)。通过将PCR产物与从含有已知螨类物种ITS拷贝的质粒中获得的产物一起进行电泳来鉴定螨类。多重PCR检测快速、可重复,灵敏度与测序相当。它被用于鉴定来自世界各地醋栗上的螨类标本。它还从红醋栗上的螨类中鉴定出一种在已知的瘿螨物种中没有对应的图谱。对这种螨的序列和生态分析表明,它是一种非造瘿瘿螨的新物种。

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