Quantitative estimation of the replication kinetics of genotype 2 PRRSV strains with different levels of virulence in vitro

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/mu L, and the assay was linear between 10(1) and 10(8) copies/mu L. 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 9611 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. (C) 2016 Elsevier B.V. All rights reserved.