Simultaneous detection of classical PRRSV, highly pathogenic PRRSV and NADC30-like PRRSV by TaqMan probe real-time PCR

Wenbin Qiu; Kai Meng; Yanyan Liu; Yuyu Zhang; Zhao Wang; Zhi Chen; Jie Yang; Wenbo Sun; Lihui Guo; Sufang Ren; Lei Chen; Guiwen Yang; Fan Zhang; Jianli Shi; Jun Li; Yijun Du; Jiang Yu; Jiaqiang Wu

doi:10.1016/j.jviromet.2019.113774

Simultaneous detection of classical PRRSV, highly pathogenic PRRSV and NADC30-like PRRSV by TaqMan probe real-time PCR

J Virol Methods. 2019 Nov 11:282:113774. doi: 10.1016/j.jviromet.2019.113774. Online ahead of print.

Authors

Wenbin Qiu¹, Kai Meng², Yanyan Liu³, Yuyu Zhang³, Zhao Wang⁴, Zhi Chen³, Jie Yang³, Wenbo Sun³, Lihui Guo³, Sufang Ren³, Lei Chen⁵, Guiwen Yang⁵, Fan Zhang⁵, Jianli Shi³, Jun Li³, Yijun Du³, Jiang Yu⁶, Jiaqiang Wu⁷

Affiliations

¹ Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China; School of Life Sciences, Shandong Normal University, Jinan, Jinan, 250014, China.
² Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China; Shandong Key Laboratory of Poultry Diseases Diagnosis and Immunology, Poultry Breeding Engineering Technology Center of Shandong Province, Institute of Poultry Science, Shandong Academy of Agricultural Sciences, Jinan, 250023, China.
³ Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China.
⁴ China Institute of Veterinary Drug Control, 8 Nandajie, Zhongguancun, Haidian, Beijing, 100081, China.
⁵ School of Life Sciences, Shandong Normal University, Jinan, Jinan, 250014, China.
⁶ Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China. Electronic address: yujiang_2213@163.com.
⁷ Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China; School of Life Sciences, Shandong Normal University, Jinan, Jinan, 250014, China; Shandong Key Laboratory of Poultry Diseases Diagnosis and Immunology, Poultry Breeding Engineering Technology Center of Shandong Province, Institute of Poultry Science, Shandong Academy of Agricultural Sciences, Jinan, 250023, China. Electronic address: wujiaqiang2000@sina.com.

PMID: 31726113
DOI: 10.1016/j.jviromet.2019.113774

Abstract

Porcine Reproductive and Respiratory Syndrome (PRRS), an acute infectious disease caused by the porcine reproductive and respiratory syndrome virus (PRRSV), is one of the most devastating diseases affecting the global swine industry. In order to establish a multiplex real-time PCR method for the simultaneous detection of the classical PRRSV (C-PRRSV) strain, the highly pathogenic PRRSV (HP-PRRSV) strain and NADC30-like PRRSV (NL-PRRSV) strain, we designed specific primers and TaqMan fluorescent probes based on the Nsp2 target gene sequence of these three different PRRSV strains, and designed American-type PRRSV (PRRSV-U) special primers and probes based on the relatively conserved target gene sequence of ORF7. The method established in this study can quickly and accurately detect and differentiate three types of strains of clinical tissue samples, respectively. This method plays a key role in the rapid diagnosis and determination of PRRSV.

Keywords: Classical PRRSV; Highly pathogenic PRRSV; NADC30-like PRRSV; Probe real-time PCR; TaqMan.