用于鉴定马铃薯孢囊线虫和烟草孢囊线虫的多重实时荧光定量PCR检测方法

Multiplex Real-Time PCR Assays for the Identification of the Potato Cyst and Tobacco Cyst Nematodes.

作者信息

Nakhla Mark K, Owens Kristina J, Li Wenbin, Wei Gang, Skantar Andrea M, Levy Laurene

机构信息

National Plant Germplasm and Biotechnology Laboratory, United States Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS) PPQ-CPHST.

Nematology Laboratory, USDA Agricultural Research Service.

出版信息

Plant Dis. 2010 Aug;94(8):959-965. doi: 10.1094/PDIS-94-8-0959.

DOI:10.1094/PDIS-94-8-0959

PMID:30743486

Abstract

TaqMan primer-probe sets were developed for the detection and identification of potato cyst nematodes (PCNs) Globodera pallida and G. rostochiensis using two-tube, multiplex real-time polymerase chain reaction (PCR). One tube contained a primer-probe set specific for G. pallida (pale potato cyst nematode) multiplexed with another primer-probe set specific for G. rostochiensis (golden potato cyst nematode). A second tube consisted of the G. pallida-specific primer-probe set multiplexed with a primer-probe set specific for G. tabacum (the morphologically similar tobacco cyst nematode). This internal transcribed spacer rDNA-based system was specific for the Globodera spp. of interest and successfully identified several populations of PCN. This rapid, sensitive, and specific quantitative PCR assay presents a useful tool for PCN regulatory response and management programs.

摘要

开发了TaqMan引物-探针组，用于使用两管多重实时聚合酶链反应（PCR）检测和鉴定马铃薯胞囊线虫（PCN），即苍白球孢囊线虫和罗斯托克球孢囊线虫。一管包含对苍白球孢囊线虫（淡色马铃薯胞囊线虫）特异的引物-探针组，与对罗斯托克球孢囊线虫（金色马铃薯胞囊线虫）特异的另一引物-探针组进行多重反应。第二管由对苍白球孢囊线虫特异的引物-探针组与对烟草胞囊线虫（形态相似的烟草胞囊线虫）特异的引物-探针组进行多重反应组成。这种基于内转录间隔区rDNA的系统对感兴趣的球孢囊线虫属特异，并成功鉴定了几个PCN种群。这种快速、灵敏且特异的定量PCR检测方法为PCN监管应对和管理计划提供了一种有用的工具。