Two new real-time quantitative reverse transcription polymerase chain reaction assays with unique target sites for the specific and sensitive detection of lineages 1 and 2 West Nile virus strains

被引：100

作者：

Eiden, Martin

Vina-Rodriguez, Ariel

Hoffmann, Bernd ^{[2
]}

Ziegler, Ute

Groschup, Martin H. ^{[1
]}

机构：

[1] Friedrich Loeffler Inst, Fed Res Inst Anim Hlth, Inst Novel & Emerging Infect Dis, Sildufer 10, D-17493 Greifswald, Germany

[2] Friedrich Loeffler Inst, Inst Diagnost Virol, D-17493 Greifswald, Germany

来源：

JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION | 2010年 / 22卷 / 05期

关键词：

Real-time quantitative reverse transcription polymerase chain reaction; West Nile virus; PCR ASSAYS; FLAVIVIRUSES; BIRDS;

D O I：

10.1177/104063871002200515

中图分类号：

S85 [动物医学（兽医学）];

学科分类号：

090604 [动物药学];

摘要：

Two novel 1-step real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays for the simultaneous detection of West Nile virus (WNV) lineage 1 and 2 strains were developed. Primers and the probe of assay 1 target the 5'-untranslated region (UTR), whereas the amplicon of assay 2 is located in the nonstructural region NS2A, which enables an unambiguous and independent WNV diagnosis based on 2 different amplicons. Both assays allow the detection of as few as 2-4 genome copies of WNV strains NY99, Uganda B956, Kunjin, and Sarafend (all cultured on Vero cells). A new synthetic RNA mutant of the 5'-UTR amplicon, which contains 6 twist inverted base-pair changes at the probe attachment site, was used as external calibrator control.

引用

页码：748 / 753

页数：6

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