title: Quantitative estimation of the replication kinetics of genotype 2 PRRSV strains with different levels of virulence in vitro.
abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) has become an important pathogen for the swine industry, and has resulted in substantial economic losses. In 2006, highly pathogenic PRRSV (HP-PRRSV) belonging to genotype 2 was first identified in China. Here, the replication kinetics of genotype 2 PRRSV strains were estimated in vitro in MARC-145 cells and porcine alveolar macrophages (PAMs) using a TaqMan-based real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. The lower limit of detection was 10 copies/μL, and the assay was linear between 10(1) and 10(8) copies/μL. The intra-assay coefficients of variation were 0.81-1.36%, and the inter-assay coefficients of variation were 1.77-2.56%. Compared to the low pathogenicity CH-1a-F45 strain, the viral loads of the highly pathogenic HuN4-F45 strain were 10(0.5)-10(1.05) and 10(0.84)-10(1.35) times greater in MARC-145 cells and PAMs, respectively from 12 to 96h after infection (P<0.01). This study is the first to demonstrate that the HuN4-F45 strain replicated at higher levels than CH-1a-F45 in MARC-145 cells and PAMs, suggesting that HuN4-F45 has more robust virus amplification efficiency than CH-1a-F45 in vitro.
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China; Xinyang Animal Disease Prevention and Control Engineering Research Center, Xinyang College of Agriculture and Forestry, Xinyang 464000, PR China.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Dong
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
authors.forename: Wenda
authors.initials: W
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
authors.forename: Jianan
authors.initials: J
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
authors.forename: Huiqiang
authors.initials: H
authors.lastname: Wen
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
authors.forename: Shujie
authors.initials: S
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Tian
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. Electronic address: cai139@hvri.ac.cn.
authors.forename: Xuehui
authors.initials: X
authors.lastname: Cai
mesh_terms: D000818:Animals; D005838:Genotype; D007700:Kinetics; D057230:Limit of Detection; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity* / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology; D019562:Viral Load; D014774:Virulence; D014779:Virus Replication
keywords: PRRSV; RT-qPCR; Replication kinetics; Viral loads