Duplex Real-Time RT-PCR Assays for the Detection and Typing of Epizootic Haemorrhagic Disease Virus

被引：20

作者：

Viarouge, Cyril ^{[1
]}

Breard, Emmanuel ^{[1
]}

Zientara, Stephan ^{[1
]}

Vitour, Damien ^{[1
]}

Sailleau, Corinne ^{[1
]}

机构：

[1] ANSES INRA ENVA UPEC, UMR Virol 1161, F-94700 Maisons Alfort, France

来源：

PLOS ONE | 2015年 / 10卷 / 07期

关键词：

POLYMERASE-CHAIN-REACTION; BLUETONGUE-VIRUS; NUCLEIC-ACID; SEROTYPES; CATTLE; ASSIGNMENT; PROTEINS; SEGMENTS; GENOME; ISLAND;

D O I：

10.1371/journal.pone.0132540

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Epizootic haemorrhagic disease virus (EHDV) may cause severe clinical episodes in some species of deer and sometimes in cattle. Laboratory diagnosis provides a basis for the design and timely implementation of disease control measures. There are seven distinct EHDV serotypes, VP2 coding segment 2 being the target for serotype specificity. This paper reports the development and validation of eight duplex real-time RT-PCR assays to simultaneously amplify the EHDV target (S9 for the pan-EHDV real-time RT-PCR assay and S2 for the serotyping assays) and endogenous control gene Beta-actin. Analytical and diagnostic sensitivity and specificity, inter-and intra-assay variation and efficiency were evaluated for each assay. All were shown to be highly specific and sensitive.

引用

页数：11