Detection of Mycobacterium kansasii using a combination of loop-mediated isothermal amplification (LAMP) and lateral flo
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ORIGINAL ARTICLE
Detection of Mycobacterium kansasii using a combination of loop-mediated isothermal amplification (LAMP) and lateral flow biosensors Chuang Chen 1 & Jia Lu 1 & Bo Long 1 & Zhengyuan Rao 1 & Yuan Gao 1 & Weina Wang 1 & Wenfeng Gao 1 & Jun Yang 1 & Shu Zhang 1 Received: 30 December 2019 / Revised: 9 August 2020 / Accepted: 28 August 2020 # The Author(s) 2020
Abstract 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. Keywords Mycobacterium kansasii (M. kansasii) . Loop-mediated isothermal amplification (LAMP) . Lateral flow biosensor (LFB)
Introduction Nontuberculous mycobacteria (NTM) comprise most mycobacteria except the Mycobacterium tuberculosis complex (MTBC) and Mycobacterium leprae. With an increase in the number of patients suffering from immune injury (HIV infection, organ transplant patients, etc.) (Griffith et al. 2007), the number of NTM diseases has increased significantly (Wassilew et al. 2016). NTM are widely distributed in the environment, mainly in soil and water, and are classified into fast-growing and slow-growing mycobacteria. Mycobacterium kansasii, an opportunistic pathogen, is a slow-growing NTM. Patients with chronic lung disease, mycobacterial disease, malignancy, and Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10123-020-00143-z) contains supplementary material, which is available to authorized users. * Shu Zhang [email protected] 1
Sichuan Center for Disease Control and Prevention, Chengdu 610041, Sichuan Province, China
alcoholism have a higher risk of developing diseases caused by M. kansasii (Kim et al. 2019; Hirashima et al. 2014; Theodore and Charles 2002; Won-Jung et al. 2002). In China, M. kansasii is the third most common slow-growing NTM to cause clinical disease (Fang et al. 2019; Tan et al. 2018). As one of the most important pathogenic nontuberculous mycobacteria, the clinical symptoms of M. kansasii infection are similar to those seen in patients with tuberculosis and other nontuberculous mycobacterial i
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