TY - JOUR
T1 - 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
AU - Chai, Zheng
AU - Ma, Wenjun
AU - Fu, Fang
AU - Lang, Yuekun
AU - Wang, Wei
AU - Tong, Guangzhi
AU - Liu, Qinfang
AU - Cai, Xuehui
AU - Li, Xi
N1 - Funding Information:
This study was supported by grants from Major International Scientific Cooperation Projects in the National Animal Zoonoses Prevention and Control of Key Technologies (2010DFB33920).
PY - 2013/2
Y1 - 2013/2
N2 - 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.
AB - 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.
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U2 - 10.1007/s00705-012-1504-7
DO - 10.1007/s00705-012-1504-7
M3 - Article
C2 - 23070137
AN - SCOPUS:84873123487
SN - 0304-8608
VL - 158
SP - 407
EP - 415
JO - Archives of Virology
JF - Archives of Virology
IS - 2
ER -