Proportions of Mycobacterium massiliense and Mycobacterium bolletii Strains among Korean Mycobacterium chelonae-Mycobacterium abscessus Group Isolates

Abstract

Korean isolates of the Mycobacterium chelonae-Mycobacterium abscessus group, which had been isolated from two different hospitals in South Korea, were identified by PCR restriction analysis (PRA) and comparative sequence analysis of 16S rRNA genes, rpoB, and hsp65 to evaluate the proportion of four closely related species (M. chelonae, M. abscessus, M. massiliense, and M. bolletii). Of the 144 rapidly growing mycobacterial strains tested, 127 strains (88.2%) belonged to the M. chelonae-M. abscessus group. In this group, M. chelonae, M. abscessus, M. massiliense, and M. bolletii accounted for 0.8% (n ‫؍‬ 1), 51.2% (n ‫؍‬ 65), 46.5% (n ‫؍‬ 59), and 1.6% (n ‫؍‬ 2), respectively. Two isolates which showed discordant results, M. massiliense by rpoB sequence analysis and M. abscessus by hsp65 sequence analysis, were finally identified as M. massiliense based on the additional analysis of sodA and the 16S-23S internal transcribed spacer. M. abscessus group I isolates previously identified by hsp65 PRA were all found to be M. abscessus, whereas group II isolates were further identified as M. massiliense or M. bolletii by sequencing of rpoB and hsp65. Smooth, rough, or mixed colonies of both M. abscessus and M. massiliense isolates were observed. M. massiliense strains that were highly resistant to clarithromycin had a point mutation at the adenine at position 2058 (A 2058 ) or 2059 (A 2059 ) in the peptidyltransferase region of the 23S rRNA gene.Rapidly growing mycobacteria (RGM), which include members of the Mycobacterium fortuitum complex, the M. chelonae complex, and the M. smegmatis complex, are defined as mycobacteria that grow and form visible colonies on solid agar media within 7 days. Because of their low virulence, they usually cause respiratory tract or disseminated infections in persons with predisposing factors or who are immunosuppressed (27,29,32). However, recently, infections in immunocompetent persons have been reported more frequently. These cases are soft-tissue infections associated with trauma and injection and epidemics or pseudoepidemics that have occurred in hospitals (6, 12). Ninety-five percent of soft-tissue RGM infections are caused by members of the M. chelonae complex (7), which comprises M. chelonae, M. abscessus, and M. immunogenum. However, the situation is complex for the clinical microbiologist who must rapidly and correctly identify isolates because there are reports of heterogeneity in M. abscessus isolates. Two M. abscessus groups (groups I and II) were reported on the basis of hsp65 PCR-restriction fragment length polymorphism (RFLP) (11) and sequence analysis (21) of hsp65. High heterogeneity of M. abscessus isolates was also shown by rpoB analysis (2). Meanwhile, new RGM species were reported: M. massiliense (4) and M. bolletii (1). These were very closely related to M. abscessus and M. chelonae but showed different susceptibility to clarithromycin, and their pathogenic potentials were shown by infections in immunocompetent and immunocompromised hosts (13,19).RGM infections are a...

Identification of Cutaneous Mycobacterium massiliense Infections Associated with Repeated Surgical Procedures

“…The rate of isolation of M. massiliense from infected patients in Korea is much higher than that in recent reports 6,9 . An epidemic occurred in Korea associated with intramuscular injection of antimicrobial agents, and the causative microorganism was finally identified as M. massiliense 8 .…”

“…abscessus 2,7,8 ; therefore, 16S rRNA gene sequencing cannot discriminate between these species. M. massiliense can be differentiated from M. abscessus by sequencing of the rpoB, hsp65, sodA, and recA genes, or the 16S-23S rRNA internal transcribed spacer sequence [6][7][8][9] .…”

“…Since the early 1990s, genotype analysis, especially PCR sequencing, has greatly contributed to the precise and rapid identification of mycobacterial species 8 . Partial 16S rRNA gene sequence analysis is the most widely used method for identifying nontuberculous mycobacteria 9,12 . However, the 16S rRNA gene sequence of M. massiliense is identical to that of M. chelonae/M.…”

“…M. massiliense is very closely related to M. abscessus and M. chelonae 7 . It has been isolated from a patient with pneumonia, a patient with pacemaker pocket infection, an outbreak associated with intramuscular injections of antimicrobial agents, and outbreaks associated with laparoscopic surgeries and cosmetic procedures 2,5,[7][8][9][10] . Here we report the case of a cutaneous M. massiliense infection associated with a surgical procedure in an immunocompetent patient.…”

Investigation of the population structure of Mycobacterium abscessus complex strains using 17-locus variable number tandem repeat typing and the further distinction of Mycobacterium massiliense hsp65 genotypes

“…The frequency of M. massiliense isolation appears to vary regionally (Harada et al, 2012;Kim et al, 2008;van Ingen et al, 2009;Zelazny et al, 2009). Recently, it was further reported that M. massiliense in Korean patients could be subdivided into two genotypes based on hsp65 sequence analysis (Kim et al, 2012).…”