During the global COVID-19 pandemic, isothermal amplification methods (IAM) began garnering more attention as a simple, cost-effective alternative to PCR for point_of_care testing. Loop-mediated isothermal amplification (LAMP) assays have been developed for detection of various viruses and bacteria from patient samples, including SARS_CoV_2. Rapidity and reliability make LAMP assays invaluable in monitoring clinical pathogens in a public health laboratory setting.
Like RT-PCR, the addition of a reverse transcriptase (RT) is necessary to initiate LAMP detection of RNA targets, including the genome of SARS-CoV-2. Historically, the avian myeloblastosis virus (AMV)-RT was widely used for RT-LAMP. Here, Moloney Murine Leukemia Virus (M-MuLV)-RT has engineered to possess the improved thermostability, increased amplification efficiency, a reduction in RNase H activity, and a lack of endonuclease activity. These properties of the engineered M-MuLV-RT have been evaluated in RT_LAMP assays for the detection of SARS_CoV_2 in clinical specimens.