TY - JOUR KW - next generation sequencing KW - molecular epidemiology KW - modified live vaccine AU - L Schneider-Bühl AU - E Hiller AU - T Frey AU - V Akimkin AU - M Hoferer AU - R Sting AB - Porcine reproductive and respiratory syndrome virus (PRRSV) causes one of the economically most important diseases in pig farming. In view of the particularly high level of genetic variation of this virus, molecular epidemiological methods are very well suited for the characterisation of virus strains. In our study, we present the results of PRRSV testing using RT-qPCR and following ORF5, ORF4–ORF6 and whole genome sequencing (WGS) in two pig farms, connected by purchase and both suffering from PRRSV outbreaks. Analyses based on ORF5 sequences detected only a field virus strain in farm A, despite vaccination with a modified live vaccine (MLV). In farm B, the MLV used in farm A and a field virus strain could be determined. However, the ORF4–ORF6 sequences confirmed the results from farm A, but revealed two different field virus strains in farm B. This phenomenon could be resolved using next generation sequencing (NGS) technologies for WGS analyses of two representative virus strains originating from farm B. WGS revealed two closely related field virus strains with a nucleotide sequence identity of 97.72%. One strain was a recombinant PRRSV strain, apparently created from the MLV used in farm A and a field virus strain. The fact that only the ORF5 region originated from the MLV explains the differences in the sequences of ORF5 compared to ORF4–ORF6 and WGS. Our study shows that ORF-based molecular characterisation of PRRSV strains might be deceptive due to mosaic recombination events requiring WGS to accurately reveal the molecular relationships between virus strains. BT - Berliner und Münchener Tierärztliche Wochenschrift CY - Hannover DA - 10/2020 DO - 10.2376/1439-0299-2020-19 LA - Englisch N2 - Porcine reproductive and respiratory syndrome virus (PRRSV) causes one of the economically most important diseases in pig farming. In view of the particularly high level of genetic variation of this virus, molecular epidemiological methods are very well suited for the characterisation of virus strains. In our study, we present the results of PRRSV testing using RT-qPCR and following ORF5, ORF4–ORF6 and whole genome sequencing (WGS) in two pig farms, connected by purchase and both suffering from PRRSV outbreaks. Analyses based on ORF5 sequences detected only a field virus strain in farm A, despite vaccination with a modified live vaccine (MLV). In farm B, the MLV used in farm A and a field virus strain could be determined. However, the ORF4–ORF6 sequences confirmed the results from farm A, but revealed two different field virus strains in farm B. This phenomenon could be resolved using next generation sequencing (NGS) technologies for WGS analyses of two representative virus strains originating from farm B. WGS revealed two closely related field virus strains with a nucleotide sequence identity of 97.72%. One strain was a recombinant PRRSV strain, apparently created from the MLV used in farm A and a field virus strain. The fact that only the ORF5 region originated from the MLV explains the differences in the sequences of ORF5 compared to ORF4–ORF6 and WGS. Our study shows that ORF-based molecular characterisation of PRRSV strains might be deceptive due to mosaic recombination events requiring WGS to accurately reveal the molecular relationships between virus strains. PB - Schlütersche Verlagsgesellschaft mbH & Co. KG PP - Hannover PY - 2020 T1 - Comparative ORF and whole genome sequencing analysis of the porcine reproductive and respiratory syndrome virus (PRRSV) in routine samples reveal a recombinant virus strain T2 - Berliner und Münchener Tierärztliche Wochenschrift TI - Comparative ORF and whole genome sequencing analysis of the porcine reproductive and respiratory syndrome virus (PRRSV) in routine samples reveal a recombinant virus strain TT - ORF- und Gesamtgenomsequenzierung des porcinen reproduktiven und respiratorischen Syndrom-Virus (PRRSV) in Routineproben führen zum Nachweis eines rekombinanten Virusstammes VL - 133 SN - 1439-0299 ER -