Development and evaluation of a real-time RT-PCR assay for Mammarenavirus guanaritoense virus detection

Mammarenavirus guanaritoense (formerly Guanarito virus, GTOV) is an important etiological agent of an emergent disease with a high mortality rate, namely Venezuelan hemorrhagic fever (VHF). Traditionally, VHF is an important public health problem and is a mandatory reportable disease. Moreover, global interconnection has increased the probability of worldwide spread of GTOV. Therefore, national healthcare organizations need tools for the management of GTOV-associated risks. These tools should definitely include diagnostic kits based on real-time reverse transcription PCR (RT-PCR) because it is the most suitable method for the diagnosis of viral hemorrhagic fever diseases. Here, we describe a real-time RT-PCR assay for the detection of GTOV. This assay was developed and evaluated using armored positive control particles (ARC). The LOD of the assay is in the range 1–2.5 × 102 RNA copies/mL on the CFX96 PCR plate machine and is in the range 1–2.0 × 103 RNA copies/mL on the Rotor-Gene Q PCR rotary machine. Our assay was evaluated using positive GTOV samples from the collection of the Instituto Nacional de Higiene “Rafael Rangel”, Caracas, Venezuela (INHRR). The assay provides a fast and sensitive tool for GTOV detection. The high specificity and sensitivity of the assay make it useful for clinical and epidemiological investigations in the field of VHF and their etiological agents.