Comparison of an Automated Repetitive Sequence-based PCR Microbial Typing System with IS6110-Restriction Fragment Length Polymorphism for Epidemiologic Investigation of Clinical Mycobacterium tuberculosis Isolates in Korea

Abstract

Tuberculosis remains a severe public health problem worldwide. Presently, genotyping is used for conducting epidemiologic and clinical studies on tuberculosis cases. We evaluated the efficacy of the repetitive sequence-based PCR (rep-PCR)-based DiversiLab™ system (bioMérieux, France) over the IS6110-restriction fragment length polymorphism analysis for detecting Mycobacterium tuberculosis. In all, 89 clinical M. tuberculosis isolates collected nationwide from Korea were used. The DiversiLab system allocated the 89 isolates to 8 groups with 1 unique isolate when a similarity level of 95% was applied. Seventy-six isolates of the Beijing family and 13 isolates of non-Beijing family strains were irregularly distributed regardless of rep-PCR groups. The DiversiLab system generated a rapid, sensitive, and standardized result. It can be used to conduct molecular epidemiologic studies to identify clinical M. tuberculosis isolates in Korea.

Comparison of a Semiautomated Commercial Repetitive-Sequence-Based PCR Method with Spoligotyping, 24-Locus Mycobacterial Interspersed Repetitive-Unit–Variable-Number Tandem-Repeat Typing, and Restriction Fragment Length Polymorphism-Based Analysis of IS 6110 for Mycobacterium tuberculosis Typing

“…1). Jang et al (27) previously suggested that DL discriminates efficiently among Beijing family strains with near-identical IS6110-RFLP patterns, in contrast with our data which rather suggest that DL is not reliable for analysis of Beijing strains. Finally, regarding the six epidemiologically unlinked T2-T3 (SIT73) isolates S46 to S51 (Table 1), the high similarity (97%) observed within the single cluster as found with DL was not consistent with the significant differences observed in their MIRU codes and especially their RFLP patterns (Fig.…”

“…The DiversiLab (DL) microbial typing system (bioMérieux, Marcy l'Etoile, France) (18) consists of a semiautomated highly standardized rep-PCR (18,19). Studies reporting on DL performance in the analysis of mycobacteria are scarce and have been done with specific aims, e.g., rapid genotyping of nontuberculosis mycobacteria (20)(21)(22)(23)(24), analysis of M. tuberculosis microevolution within a patient (25), monitoring of TB outbreaks (1,3,26), and comparison of DL to other molecular techniques for M. tuberculosis typing (1,19,27,28).…”

Methodological and Clinical Aspects of the Molecular Epidemiology of Mycobacterium tuberculosis and Other Mycobacteria

“…Data generated with this method showed high concordance with those generated by IS6110-RFLP and IS1245-RFLP typing for M. tuberculosis and MAC strains, respectively (410). This method was also able to discriminate between M. tuberculosis Beijing strains whose IS6110-RFLP patterns were nearly identical (411). More recently, automated rep-PCR was applied for typing of M. abscessus and compared with the PFGE method.…”

Molecular characterization of Mycobacterium tuberculosis and Mycobacterium bovis isolates by Enterobacterial Repetitive Intergenic Consensus-PCR

“…In the study conducted by Noguti et al (2010), there were two isolates. Cangelosi et al (2004), Ashworth et al (2008) and Jang et al (2011) evaluated the utility of a commercially available rep-PCR System, initially developed for genotyping Streptococcus pneumoniae, Staphylococcus aureus, Enterobacteriaceae and others microorganism, to genotype M. tuberculosis and Mycobacterium avium Complex and found the system to be highly discriminative.…”