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Porcine reproductive and respiratory syndrome (PRRS) viruses are recognized as possessing a high degree of genetic and antigenic variability. Viral diversity has led to questions regarding the association of virus mutation and persistent infection in the host and raised concerns vis-a-vis protective immunity, the ability of diagnostic assays to detect novel variants, and the possible emergence of virulent strains. The purpose of this study was to describe on-going changes in PRRS virus during replication in pigs under experimental conditions. Animals were inoculated with a plaque-cloned virus derived from VR-2332, the North American PRRS virus prototype. Three independent lines of in vivo replication were maintained for 367 days by pig-to-pig passage of virus at 60 day intervals. A total of 315 plaque-cloned viruses were recovered from 21 pigs over the 367 day observation period and compared to the original plaque-cloned virus by virus neutralization assay, monoclonal antibody analysis, and sequencing of open reading frames (ORFs) lb (replicase), 5 (major envelope protein), and 7 (nucleocapsid) of the genome. Variants were detectable by day 7 post inoculation and multiple variants were present concurrently in every pig sampled over the observation period. Sequence analysis showed ORFs lb and 7 to be highly conserved. In contrast, sequencing of ORF

Evolution of porcine reproductive and respiratory syndrome (PRRS) virus during sequential pig passages