A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China

Zheng Chai, Wenjun Ma, Fang Fu, Yuekun Lang, Wei Wang, Guangzhi Tong, Qinfang Liu, Xuehui Cai, Xi Li

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Abstract

SYBR Green coupled to melting curve analysis has been suggested to detect RNA viruses showing high genomic variability. Here, a SYBR Green-based real-time RT-PCR assay was developed for simultaneous detection and differentiation of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) and classical type 2 PRRSV (C-PRRSV). The different strains were identified by their distinctive melting temperatures: 82. 98 ± 0. 25 °C and 85. 95 ± 0. 24 °C for HP-PRRSVs or 82. 74 ± 0. 26 °C for C-PRRSVs. Specificity was tested using nine other viral and bacterial pathogens of swine. The detection limit was 1 TCID50 for HP- or C-PRRSV. Furthermore, the detection results for samples from an animal trial with HP- or C-PRRSV infections showed that the SYBR Green-based real-time RT-PCR was more sensitive than the conventional RT-PCR. Additionally, an analysis of 319 field samples from North China, Central China and Northeast China showed that HP- and C-PRRSVs co-circulated in pig herds. Thus, the SYBR Green-based real-time RT-PCR, which can be performed within one hour, is a rapid, sensitive and low-cost diagnostic tool for rapid differential detection and routine surveillance of HP- and classical type 2 PRRSVs in China.

Original languageEnglish (US)
Pages (from-to)407-415
Number of pages9
JournalArchives of Virology
Volume158
Issue number2
DOIs
StatePublished - Jan 1 2013

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Porcine respiratory and reproductive syndrome virus
Real-Time Polymerase Chain Reaction
China
Freezing
Swine
RNA Viruses
Limit of Detection
Costs and Cost Analysis
Polymerase Chain Reaction
Temperature

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A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China. / Chai, Zheng; Ma, Wenjun; Fu, Fang; Lang, Yuekun; Wang, Wei; Tong, Guangzhi; Liu, Qinfang; Cai, Xuehui; Li, Xi.

In: Archives of Virology, Vol. 158, No. 2, 01.01.2013, p. 407-415.

Research output: Contribution to journalArticle

Chai, Zheng ; Ma, Wenjun ; Fu, Fang ; Lang, Yuekun ; Wang, Wei ; Tong, Guangzhi ; Liu, Qinfang ; Cai, Xuehui ; Li, Xi. / A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China. In: Archives of Virology. 2013 ; Vol. 158, No. 2. pp. 407-415.
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