Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 161, Issue 2, Pages 192-198Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2009.06.007
Keywords
Classical US PRRSV; Highly pathogenic US PRRSV; Differential detection; Duplex real-time RT-PCR; Minor groove binder probe
Funding
- Ministry of Science and Technology of China [2006BAD06A12, 2007BAD861303]
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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 3.2 TCID50/ml or 38 RNAcopies/mu l for C-US-PRRSV and 0.4TCID(50)/ml or 14 RNA copies/mu l 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 2 h. This duplex real-time RT-PCR assay is a promising tool for rapid differential detection and epidemiology of US PRRSV in China. (C) 2009 Elsevier B.V. All rights reserved.
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