Development of real-time RT-PCR assays for two viruses infecting pome fruit

E Beaver-Kanuya; S A Szostek; S J Harper

doi:10.1016/j.jviromet.2018.12.008

Development of real-time RT-PCR assays for two viruses infecting pome fruit

J Virol Methods. 2019 Apr:266:25-29. doi: 10.1016/j.jviromet.2018.12.008. Epub 2019 Jan 14.

Authors

E Beaver-Kanuya¹, S A Szostek², S J Harper²

Affiliations

¹ Department of Plant Pathology, Washington State University, Prosser, WA, 99350, United States. Electronic address: eunice.kanuya@wsu.edu.
² Department of Plant Pathology, Washington State University, Prosser, WA, 99350, United States.

PMID: 30650343
DOI: 10.1016/j.jviromet.2018.12.008

Abstract

Apple stem grooving virus (ASGV) and Apple green crinkle-associated virus (AGCaV) negatively impact production, maintenance, and distribution of apples and other Malus species world-wide. Due to the increasing diversity of isolates found by high-throughput sequencing, we have developed real-time RT-qPCR assays for these two viruses. Primers and probes were designed against alignments of representative extant sequences from around the world, and reaction conditions optimized for sensitivity and specificity. Assays were validated against a panel of virus isolates, and compared to extant endpoint RT-PCR and ELISA assays. The new real-time RT-qPCR assays showed greater detection sensitivity than extant assays and were able to detect their target viruses from different host tissues.

Keywords: AGCaV; ASGV; Malus; real-time RT-qPCR.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

DNA Primers / genetics
Malus / virology*
Plant Diseases / virology*
Plant Viruses / isolation & purification*
RNA, Viral / isolation & purification*
Reverse Transcriptase Polymerase Chain Reaction / methods*
Sensitivity and Specificity

Substances

DNA Primers
RNA, Viral