Multiplex Pathogen Screening in Swine

Nucleic acids were extracted from moMΦ using a MagMax Core kit and MagMax96 extractor (Thermo Fisher) according to the manufacturer’s instructions; samples were kept at −20 °C until analyzed. The presence of PRRSV or M. hyopneumoniae genome was subsequently screened using commercial real-time PCR kits, LSI VetMAX™ PRRSV EU/NA and VetMAX™-Plus qPCR Master Mix (both Thermo Fisher Scientific), respectively. The presence of PCV2 was instead evaluated by qualitative real-time PCR, as we previously described [44 (link)], using forward primer 5′-TGGCCCGCAGTATTCTGATT-3′ and reverse primer 5′-CAGCTGGGACAGCAGTTGAG-3′ [45 (link)]. Samples with Ct values less than 40 were considered positive [44 (link)].