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Abstract
Mycobacterium abscessus (M. abscessus) and Mycobacterium massiliense (M. massiliense) are major pathogens that cause post-surgical wound infection and chronic pulmonary disease. Although they are closely related subspecies of M. abscessus complex, their infections are associated with different drug-resistance and cure rate. In the present study, a loop-mediated isothermal amplification (LAMP) coupled with lateral flow dipstick (LFD) method was developed to simultaneous detect M. abscessus and M. massiliense, via specific erm(41) gene. The amplification was carried out at 65 °C for only 60 min, and the results could be visualized on a lateral flow strip. Positive results only occurred in M. abscessus and M. massiliense, no cross-reaction with other mycobacterial species was observed. Therefore, the cost-effective MABC (M. abscessus complex)–LAMP–LFD method developed here was able to correct the diagnose of M. abscessus and M. massiliense infection in a short time. Thus, this method could be used to guide clinicians in treatment of M. abscessus group infections.
Highlights
Growing mycobacteria (RGM) are ubiquitous environmental microorganisms (Brown-Elliott and Wallace 2002) and the prevalence of pulmonary infection due to Rapidly growing mycobacteria (RGM) is increasing worldwide
Presence of the erm(41) gene in clinical isolates The novel erm(41) gene was found to be unique to the M. abscessus complex
We described a simple, robust, accurate, rapid and cost-effective loop-mediated isothermal amplification (LAMP)-based method to detect and distinguish M. abscessus and M. massiliense from clinical specimens directly
Summary
Growing mycobacteria (RGM) are ubiquitous environmental microorganisms (Brown-Elliott and Wallace 2002) and the prevalence of pulmonary infection due to RGM is increasing worldwide. Infections caused by M. abscessus complex are often difficult to treat, because these mycobacteria are intrinsically resistant to the traditional antituberculous drugs and to most currently available antimicrobial agents (Kim et al 2015). Few M. massiliense strains have shown inducible resistance to clarithromycin because of a large (274bp) fragment deletion in M. massiliense’s erm(41) gene (Kim et al 2010) To exploit this genetic difference, PCR-based assays of this conserved deletion in erm(41) have been proposed as a simple method to distinguish M. abscessus from M. massiliense, based on the fragment size of the resulting amplification product (Kim et al 2010). Targeted gene sequencing (hsp, rpoB and secA1) and PCR-based assays (erm(41)) have been used for identification of M. abscessus and M. massiliense (Shallom et al 2013; Zelazny et al 2009), these two methods are time-consuming in a routine clinical microbiology laboratory and often require instrumentation that may not be available in resource-limited settings. We conducted drug susceptibility testing (DST) on M. abscessus complex clinical isolates to confirm their drug susceptibility patterns
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