Development of Real-Time Reverse Transcriptase PCR Assays for the Detection of Punta Toro Virus and Pichinde Virus
USAMRIID Frederick United States
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Research with high biocontainment pathogens such as Rift Valley fever virus RVFV and Lassa virus LASV is expensive, potentially hazardous, and limited to select institutions. Surrogate pathogens such as Punta Toro virus PTV for RVFV infection and Pichinde virus PICV for LASV infection allow research to be performed under more permissive BSL-2 conditions. Although used as infection models, PTV and PICV have no standard real-time RT-PCR assays to detect and quantify pathogenesis. PTV is also a human pathogen, making a standardized detection assay essential for biosurveillance. Here, we developed and characterized two real-time RT-PCR assays for PICV and PTV by optimizing assay conditions and measuring the limit of detection LOD and performance in multiple clinical matrices.