Detection ofMycobacterium kansasiiusing a combination of loop-mediated isothermal amplification (LAMP) and lateral flow biosensors

Mycobacterium kansasiiis an opportunistic pathogen that causes both intrapulmonary and extrapulmonary infections. The symptoms of the pulmonary diseases caused byM. kansasiiclosely resembleMycobacterium tuberculosis. Rapid and accurate differentiation ofM. kansasiifromM. tuberculosis, as well as other mycobacteria, is crucial for developing effective therapeutics and disease treatment. In this study, we combined loop-mediated isothermal amplification (LAMP) with lateral flow biosensors (LFB) to detectM. kansasii, by targeting the species-specific sequence ofrpoB, a gene which encodes the beta subunit of bacterial RNA polymerase. The assay was validated to ensure that it was highly selective by testingM. kansasii,M. tuberculosis, other species of respiratory bacteria, and other nontuberculous mycobacteria. The detection limit of the assay was 1 fg/mu L of DNA and 50 CFU of bacilli in sputum. TheM. kansasii-LAMP-LFB assay is a fast, cheap, and accurate method for detectingM. kansasiiby constant temperature amplification and simple interpretation.