Identification and Molecular Epidemiology of Mycobacterial Isolates in Kayseri/Turkey

Abstract

Background: Early diagnosis and treatment of tuberculosis (TB) patients are the most effective ways to protect society from this disease. The objectives of this study were to evaluate the molecular epidemiology of mycobacterial strains by repetitive sequence PCR (rep-PCR) using the DiversiLab System method at Kayseri in Turkey. Methods: A total of fifteen clinically sample isolated Mycobacterial strains were identified as M. tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) by the rep-PCR method (bioMerieux), SD (Standard Diagnostics) rapid tests. Genotyping was performed by the rep-PCR method and clonal affinity was researched. Results: Nine of the strains were identified as MTBC and six strains were identified as NTM by the SD rapid test methods. Five of the strains were identified as NTM by the rep-PCR method as two M. scrofulaceum, M. chelonae, M. fortuitum and M. avium complex (MAC). By genotyping with rep-PCR a total of nine clones were identified, which included two main clones (A and B) and seven other clones (C, D, E, F, G, H, I). Conclusions: It was detected that rep-PCR test is useful method and can be used for the molecular epidemiology and for the identification of mycobacterial strains as they are rapid and practical. However, it is necessary to perform new studies for more mycobacterial strains using the methods described herein.

A novel repeat sequence-based PCR (rep-PCR) using specific repeat sequences of Mycobacterium intracellulare as a DNA fingerprinting

“…In the MAC-PD, a recurrence caused by strains having different genotypes are frequent ( Koh et al, 2017 ; Akoglu, 2018 ; Heidari et al, 2018 ; Gautam, 2022 ; Wang et al, 2022 ), requiring a more advanced approach than naïve species identification. Various studies on the efficacy of NTM treatment have employed epidemiological approaches of the intraspecies classification level using ( Englund, 2003 ; Bussone et al, 2012 ; van der Zanden et al, 2012 ; San Millán et al, 2013 ; Thomson et al, 2013 , 2014 ; Koh et al, 2017 ; Heidari et al, 2018 ; Karauz et al, 2018 ; van Weezep et al, 2019 ; Gautam, 2022 ; Zhao et al, 2022 ). Since WGS, PFGE, and VNTR require a specialized technique, labor-intensive or time-consuming approaches, several clinical studies used rep-PCR ( Cangelosi et al, 2004 ; Jagielski et al, 2016 ; Jhun et al, 2018a , b ; Shin et al, 2020a , b ; Furuuchi et al, 2022 ) for diagnosis of reinfection/relapse, recurrent after latency, evaluation of antibiotics therapy in recurrent MAC-PD, and MAC bacteremia follow-up study.…”

“…Among the technologies related to rep-PCR, a commercial kit for rep-PCR-based mycobacteria strain typing was released in 2003 and has been used in many clinical epidemiological studies on the therapeutic efficiency of MAC ( Bussone et al, 2012 ; van der Zanden et al, 2012 ; Thomson et al, 2013 , 2014 ; Koh et al, 2017 ; Jhun et al, 2018a , b ; Karauz et al, 2018 ), including M. intracellulare . However, due to the discontinuation of the product, follow-up research is impossible.…”