致死粒线虫重组酶聚合酶扩增检测方法的建立
Development of recombinase polymerase amplification assay for detection of Anguina funesta
林宇 1刘娟 1俞禄珍 2李兰 3姜一 1贺艳 1王金成 1魏亚东1
作者信息
- 1. 天津海关 天津 300456
- 2. 上海海关
- 3. 阿拉山口海关
- 折叠
摘要
致死粒线虫是一种传播扩散风险较高的外来有害生物,对畜牧业生产具有严重危害.为实现致死粒线虫的高效准确鉴定,本研究根据致死粒线虫核糖体28S rRNA基因D2/D3片段的保守区域设计特异引物,建立了致死粒线虫的RPA检测方法.结果表明:建立的RPA检测方法对致死粒线虫具有好的特异性,扩增出230bp的特异性目标片段,检测灵敏度可达1/160条线虫.该方法检测时间短、特异性好、灵敏度高,适用于致死粒线虫的快速检测.
Abstract
Anguina funesta is an invasive pest with a high risk of transmission and spread,which has ex-tremely harmful to animal husbandry production.In order to detect A.funesta accurately and effectively,specific primers were designed based on the conserved regions of the 28S rRNA-D2/D3 to develop a recom-binase polymerase amplification(RPA)assay for the detection of A.funesta.The results showed that the RPA assay proved to be specific,a fragment of 230 bp was amplified from A.funesta by specific primers.The sensitivity test showed that the RPA assay can detect 1/160 of a single nematode DNA.The results indicate the RPA assay method had advantages of rapidity,specificity,sensitivity,and suitable for rapid detection of A.funesta.
关键词
致死粒线虫/重组酶聚合酶扩增/28S/rRNA基因/鉴定Key words
Anguina funesta/recombinase polymerase amplification/28S rRNA gene/detection引用本文复制引用
出版年
2024
NETL
NSTL
万方数据