A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus

Abstract

Vesiculoviruses (VSV) are zoonotic viruses that cause vesicular stomatitis disease in cattle, horses and pigs, as well as sporadic human cases of acute febrile illness. Therefore, diagnosis of VSV infections by reliable laboratory techniques is important to allow a proper case management and implementation of strategies for the containment of virus spread. We show here a sensitive and reproducible real-time reverse transcriptase polymerase chain reaction (RT-PCR) for detection and quantification of VSV. The assay was evaluated with arthropods and serum samples obtained from horses, cattle and patients with acute febrile disease. The real-time RT-PCR amplified the Piry, Carajas, Alagoas and Indiana Vesiculovirus at a melting temperature 81.02 ± 0.8ºC, and the sensitivity of assay was estimated in 10 RNA copies/mL to the Piry Vesiculovirus. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods. Thus, this assay allows a preliminary differential diagnosis of VSV infections.

Increased Virulence of an Epidemic Strain of Vesicular Stomatitis Virus Is Associated With Interference of the Innate Response in Pigs

“…Importantly, this is the first report demonstrating the presence of viral RNA in serum and blood samples of pigs experimentally infected with VSV. This finding is consistent with previous reports describing virus RNA in cattle and horses naturally infected with VSV ( Tolardo et al, 2016 ), and horses experimentally infected with VSIV ( Howerth et al, 2006 ). In fact, VSV RNA was regularly detected in serum samples from cattle naturally infected in Mexico with a viral strain associated with lineage 1.1 (Velazquez-Salinas, unpublished data).…”