Rapid differential detection of classical and highly pathogenic North American Porcine Reproductive and Respiratory Syndrome virus in China by a duplex real-time RT-PCR.
Since the emergence of highly pathogenic North American Porcine Reproductive and Respiratory Syndrome virus (H-US-PRRSV) in 2006, the classical North American PRRSV (C-US-PRRSV) and H-US-PRRSV isolates have coexisted in Chinese swine herds. A duplex real-time RT-PCR assay using minor groove binder (MGB) probes for differential detection of the two US PRRSV isolates was developed. The specificity, sensitivity, reproducibility, and interference test of this assay were validated. The sensitivity of the assay was 3.2TCID(50)/ml or 38 RNA copies/microl for C-US-PRRSV and 0.4 TCID(50)/ml or 14 RNA copies/microl for H-US-PRRSV. Both assays were 10 times more sensitive than the current methods. A total of 302 clinical samples were tested by duplex real-time RT-PCR and conventional RT-PCR assays, and the results showed over 98.7% agreement. In addition, the new assay can be completed in less than 2h. This duplex real-time RT-PCR assay is a promising tool for rapid differential detection and epidemiology of US PRRSV in China.
Chen
Nan-Hua
NH
Veterinary Diagnostic Lab, China Animal Disease Control Center, Haidian District, Beijing, China.
Chen
Xi-Zhao
XZ
Hu
Dong-Mei
DM
Yu
Xiu-Ling
XL
Wang
Li-Lin
LL
Han
Wei
W
Wu
Jia-Jun
JJ
Cao
Zhen
Z
Wang
Chuan-Bin
CB
Zhang
Qian
Q
Wang
Bao-Yue
BY
Tian
Ke-Gong
KG
D000818:Animals; D001483:Base Sequence; D002681:China; D008969:Molecular Sequence Data; D009656:North America; D019318:Porcine Reproductive and Respiratory Syndrome / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013552:Swine
D012367:RNA, Viral