Mycobacterium kansasii is an opportunistic pathogen that causes both intrapulmonary and extrapulmonary infections. The symptoms of the pulmonary diseases caused by M. kansasii closely resemble Mycobacterium tuberculosis. Rapid and accurate differentiation of M. kansasii from M. 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 detect M. kansasii, by targeting the species-specific sequence of rpoB, a gene which encodes the β subunit of bacterial RNA polymerase. The assay was validated to ensure that it was highly selective by testing M. kansasii, M. tuberculosis, other species of respiratory bacteria, and other nontuberculous mycobacteria. The detection limit of the assay was 1 fg/μL of DNA and 50 CFU of bacilli in sputum. The M. kansasii-LAMP-LFB assay is a fast, cheap, and accurate method for detecting M. kansasii by constant temperature amplification and simple interpretation.

堪萨斯分枝杆菌是一种机会性病原体，可引起肺内和肺外感染。堪萨斯分枝杆菌引起的肺部疾病的症状与结核分枝杆菌非常相似。快速准确地将堪萨斯分枝杆菌与结核分枝杆菌以及其他分枝杆菌区分开来，对于开发有效的治疗方法和疾病治疗至关重要。在这项研究中，我们通过靶向rpoB的物种特异性序列，将环介导等温扩增 (LAMP) 与侧流生物传感器 (LFB) 相结合来检测堪萨斯分枝杆菌，一种编码细菌RNA聚合酶β亚基的基因。通过测试堪萨斯分枝杆菌、结核分枝杆菌、其他呼吸道细菌和其他非结核分枝杆菌，验证了该测定以确保其具有高度选择性。该测定的检测限为1 fg/μL 的DNA 和50 CFU 的痰中杆菌。在堪萨斯分枝杆菌-LAMP-LFB测定是一种快速，廉价和准确的用于检测方法堪萨斯分枝通过恒温扩增和简单的解释。