Genotypic characterization of five subspecies of Mycobacterium kansasii

Abstract

Different molecular typing methods including restriction fragment length polymorphism (RFLP) analysis with the major polymorphic tandem repeat (MPTR) probe and the IS1652 probe, pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphism (AFLP) analysis, and PCR restriction analysis of the hsp-65 gene (PRA) were applied to clinical and water isolates of Mycobacterium kansasii. RFLP with the MPTR probe, PRA, PFGE, and AFLP analysis revealed five homogeneous clusters which appeared to be subspecies. RFLP with the MPTR probe and PRA gave patterns specific for each cluster, whereas PFGE and AFLP analysis gave polymorphic patterns. IS1652 was present in two of the five clusters and provided polymorphic patterns for one cluster only. The two IS1652-positive clusters were Accuprobe negative (Accuprobe test; Gen-Probe Inc.), and only two other clusters were Accuprobe positive. A PCR test based on the detection of a species-specific fragment (M. Yang, B. C. Ross, and B. Dwyer, J. Clin. Microbiol. 31:2769-2772, 1993) was positive for all M. kansasii strains. This PCR test is an accurate, rapid, and specific M. kansasii identification test. No subspecies was particularly more virulent, because all clusters contained clinical strains, from AIDS patients and non-AIDS patients, and environmental strains. Mycobacterium kansasii, like M. avium and M. xenopi, is one of the major pulmonary mycobacterial pathogens. The relative incidence of disease due to these pathogens varies between geographical regions. In the United States, M. avium and M. kansasii are the most frequently isolated mycobacteria, whereas M. kansasii is rare in Australia. However, in some European countries, M. kansasii and M. xenopi infections were more common than M. avium infections before the AIDS epidemic (7). M. kansasii infections are often clustered, mainly in urban areas, as reported in studies in Texas, the Czech Republic, and other regions (7, 11). M. avium predominates in human immunodeficiency virus (HIV)-infected patients, and M. avium infection represents the most frequent disseminated infection diagnosed in these patients. However, HIV-infected patients also suffer pulmonary or disseminated infections due to M. kansasii. In some areas, for example, Switzerland, the prevalence of mycobacterial disease due to M. kansasii has been unchanged by the AIDS epidemic, which was not associated with a shift to a predominance of M. avium (7). Unlike tubercle bacilli, human-to-human transmission has never been established for infections due to other mycobacteria. These mycobacterial infections are considered to be acquired from the environment. M. kansasii has been almost exclusively recovered from tap water; other environmental samples are rarely positive (3, 6, 10, 14, 23, 31). M. kansasii was found to be able to survive in tap water for up to 12 months, whereas long-term survival in soil could not be shown (10). Because the M. kansasii isolates found in drinking-water distribution systems are not contaminants from another source but are ...

Molecular epidemiology of tuberculosis and other mycobacterial infections: main methodologies and achievements

“…Symptoms of M. kansasii infections often resemble tuberculosis caused by M. tuberculosis complex bacteria. The PGRS and MPTR sequences can be used to type M. kansasii isolates with the aid of RFLP typing on the basis of these repetitive elements as probes [97, 294–296]. IS 1652 RFLP is only suitable for a limited number of M. kansassii subspecies [296, 297].…”

“…The PGRS and MPTR sequences can be used to type M. kansasii isolates with the aid of RFLP typing on the basis of these repetitive elements as probes [97, 294–296]. IS 1652 RFLP is only suitable for a limited number of M. kansassii subspecies [296, 297]. Other typing methods are, for example, 16S‐23S spacer region typing [298]; large restriction fragment analysis [299], amplified fragment length polymorphism analysis [296]; PFGE [296], and PCR restriction analysis of the hsp‐65 gene [300].…”

Mycobacteria in drinking water distribution systems: ecology and significance for human health

“…Picardeau et al [70] found identical PFGE patterns for M. kansasii isolates from three HIV-negative patients from a single family living in the same household as for a shower isolate recovered from their home. A M. avium sputum isolate from a non-AIDS patient yielded an identical PFGE pattern as an isolate from a water sample isolated at home [73].…”

Comparative study of two typing methods,hsp65 PRA and ITS sequencing, revealed a possible evolutionary link betweenMycobacterium kansasiitype I and II isolates

“…Molecular typing methods demonstrated that M. kansasii is a heterogeneous species with several distinct subtypes (Ross et al, 1992;Alcaide et al, 1997;Iinuma et al, 1997;Picardeau et al, 1997;Richter et al, 1999;Gaafar et al, 2003;Taillard et al, 2003). PCR and restriction enzyme analysis (PRA) of the hsp-65 gene has been widely applied to studies concerning M. kansasii genotypic characterization (Ross et al, 1992;Picardeau et al, 1997;Richter et al, 1999;Taillard et al, 2003;Chimara et al, 2004). To date, such investigations have suggested that M. kansasii of PRA type I is the most prevalent type from clinical isolates worldwide and is only rarely isolated from the environment (Tortoli, 2003).…”