Molecular detection and characterisation of the porcine reproductive and respiratory syndrome virus (PRRSV) by RT-PCR and DNA sequencing.

摘要

Porcine respiratory and reproductive syndrome virus (PRRSV) causes a devastating disease in swine worldwide. The syndrome leads to reproductive failure in sows, morbidity of suckling piglets and respiratory disorders in weaned pigs. The approaches currently used for detection of the virus are based on virus isolation on porcine alveolar macrophages (RAM) or in the established cell lines, CL 2621 or Marc-145. Final identification of the virus is serologically confirmed by indirect immunoperoxidase tests using specific antisera. While virus isolation provides evidence of PRRSV infection. the procedure is complicated by the continuous preparation of RAM. Thus, a sensitive method of PRRSV detection that does not rely on cell culture techniques is needed for diagnosis of PRRS. Recently, molecular biological methods based on RT-PCR have enabled the detection of PRRSV viral RNA in blood, tissue samples and semen. There is increasing evidence that European and North American isolates might represent twodifferent genotypes. Less is known about variability within these two distinct PRRSV genotypes. The objective of this study was to apply RT-PCR and DNA sequencing for fast and sensitive detection and characterization of PRRSV in Poland.