Preliminary report on the development of a colorimetric loop-mediated isothermal amplification diagnostic assay for deer tick virus
Roy, B. G.; Ryndock, E. J.
Show abstract
Deer tick virus (DTV) is an emerging pathogen in North America. This virus can cause nervous system complications such as encephalitis in humans. Further, no data has been surmounted around long-term effects of infection from DTV patients across variable age groups. Diagnostic tools of DTV used by government laboratories are based on RT-PCR using patient serum or ticks. This paper explores the feasibility of a colorimetric loop-mediated isothermal amplification (LAMP) assay to create a point-of-care diagnostic methodology for use in field and in primary care. LAMP consists of six primers that bind to target DNA and amplifies variable length nucleotide strands that can be visualized through side reactions or via electrophoresis. First, a viable LAMP primer set, and a primer set that dimerizes and amplifies DNA regardless of compatibility were created in silico and validated in vitro. Then, a specific LAMP assay was developed. Our findings showed his method can be performed within 30 minutes and can measure with limits of detection comparable to PCR.
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