Real-time RT-PCR for Mayaro virus detection in plasma and urine

Background: Mayaro virus (MAYV) causes an acute febrile illness which can be difficult to differentiate from dengue or chikungunya. MAYV RNA can be detected in plasma during the first 3-5 days of illness, but only a single rRT-PCR has been fully evaluated in the literature.& para;& para;Objectives: To develop an rRT-PCR for MAYV and evaluate assay performance using human plasma and urine samples spiked with different MAYV strains.& para;& para;Study design: A MAYV rRT-PCR targeting a region of the 5'UTR and nsp1 gene was designed from the alignment of all complete-genome MAYV sequences to be compatible with existing laboratory protocols. The assay was evaluated using human samples spiked with six MAYV strains, including strains from each of the three genotypes.& para;& para;Results: The linear range of the MAYV rRT-PCR extended from 1.0 to 8.0 log(10) copies/mu L, and the lower limit of 95% detection was 8.2 copies/mu L. No detection was observed when the MAYV rRT-PCR was tested with genomic RNA from related arboviruses. The assay demonstrated linear amplification of all 6 MAYV strains when spiked into human plasma samples as well as 2 strains spiked into urine.& para;& para;Conclusions: We report the design and evaluation of an rRT-PCR for MAYV. Given the concern for MAYV emergence in the Americas and the few molecular tests that have been evaluated in the literature, this assay should provide a useful diagnostic for patients with an acute febrile illness.