Emergence of Mycobacterium kansasii as the Leading Mycobacterial Pathogen Isolated Over a 20-Year Period at a Midwestern Veterans Affairs Hospital

Bittner, Marvin J.; Horowitz, Edward A.; Safranek, Thomas J.; Preheim, Laurel C.

doi:10.1093/clinids/22.6.1109

Cited by 31 publications

(21 citation statements)

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“…The real-time PCR assay that we describe here is simple, rapid, and appropriate in scope for the routine mycobacteriology laboratory. It facilitates the accurate identification of commonly encountered slowly growing and rapidly growing myco- (4,38). However, given the simple design of this assay, it would be possible to replace or add primer sets to meet the individual needs of those laboratories.…”

Section: Discussionmentioning

confidence: 99%

Rapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR

2009

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The rapid identification of mycobacteria from culture is of primary importance for the administration of empirical antibiotic therapy and for the implementation of public health measures, yet there are few commercially available assays that can easily and accurately identify the mycobacteria in culture in a timely manner. Here we report on the development of a multiplex, real-time PCR assay that can identify 93% of the pathogenic mycobacteria in our laboratory in two parallel reactions. The mycobacteria identified by this assay include the Mycobacterium tuberculosis complex (MTC), the M. avium complex (MAC), the M. chelonae-M. abscessus group (MCAG), the M. fortuitum group (MFG), and M. mucogenicum. The primer targets included the 16S rRNA gene and the internal transcribed spacer. The assay was initially validated with a repository of reference strains and was subsequently tested with 314 clinical cultures identified by the AccuProbe assay or high-performance liquid chromatography. Of the 314 cultures tested, multiplex, real-time PCR produced congruent results for 99.8% of the 1,559 targets evaluated. The sensitivity and the specificity were each 99% or greater for MTC (n ‫؍‬ 96), MAC (n ‫؍‬ 97), MCAG (n ‫؍‬ 68), and M. mucogenicum (n ‫؍‬ 9) and 95% and 100%, respectively, for MFG (n ‫؍‬ 19). We conclude that this multiplex, real-time PCR assay is a useful diagnostic tool for the rapid and accurate identification of MTC and clinically relevant nontuberculous mycobacteria.Mycobacterium tuberculosis, the causative agent of tuberculosis, is among the leading infectious causes of death in developing nations (23). In resource-rich countries, where tuberculosis is not endemic, nontuberculous mycobacteria (NTM) are responsible for the majority of mycobacterial infections in both immunocompromised and immunocompetent individuals (14). The clinical gravity of mycobacterial infections necessitates rapid isolation by culture and timely identification by the appropriate diagnostic assays. Rapid identification not only serves to focus empirical antibiotic therapy and thus avoid unnecessary drug exposure but also may aid with the appropriate respiratory isolation and prevention of secondary cases (15).Few molecular methods commercially available in the United States easily and rapidly identify mycobacteria in culture. Although the AccuProbe assay by Gen-Probe (San Diego, CA) is sufficient for the identification of isolates of the Mycobacterium tuberculosis complex (MTC) and the M. avium complex (MAC) (37), it lacks probe sets specific for rapidly growing mycobacteria, such as the M. M. peregrinum, and M. alvei (11). For these isolates, other methods, such as biochemical reactions (6, 33), high-performance liquid chromatography (HPLC) (13), and DNA sequencing (17), are required for identification. To address the need for a simple and rapid molecular assay with a broader identification scope, we developed a multiplex, real-time PCR assay that can identify 93% of the pathogenic mycobacterial isolates (both slowly growing and ra...

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Section: Discussionmentioning

confidence: 99%

Rapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR

2009

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“…The relative contribution of M. kansasii to mycobacterial pneumonia varies by geographical location. One study in an area of low TB endemicity even reported that the number of M. kansasii isolates exceeded that of M. tuberculosis isolates [46]. In our study, we recruited patients infected with M. kansasii from a distinct region in The Netherlands where an unusually high frequency of infection with M. kansasii had been described [47].…”

Section: Discussionmentioning

confidence: 99%

Tuberculin Skin Testing and In Vitro T Cell Responses to ESAT‐6 and Culture Filtrate Protein 10 after Infection withMycobacterium marinumorM. kansasii

et al. 2002

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T cell responses to ESAT-6 and culture filtrate protein 10 (CFP-10), antigens expressed by Mycobacterium tuberculosis but not by M. bovis bacille Calmette-Guérin (BCG), were found to discriminate reliably between infection with M. tuberculosis and BCG vaccination. Because the esat-6 and cfp-10 genes occur in M. kansasii and M. marinum, T cell responses to ESAT-6 and CFP-10 were investigated in patients infected with M. kansasii or M. marinum, persons intensively exposed to environmental mycobacteria, and unexposed control subjects. Tuberculin skin tests were performed, and peripheral blood mononuclear cells were cocultured with ESAT-6, CFP-10, peptide mixtures of ESAT-6 and CFP-10, and control antigens. When enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot assay (ELISPOT) were used to measure interferon-gamma production, most M. kansasii- or M. marinum-infected patients and several persons exposed to environmental mycobacteria were found to respond to ESAT-6 and/or CFP-10. ELISA and ELISPOT yielded comparable results, as did whole antigen and peptides (P<.0001). These results may be relevant for the development of novel assays for diagnosis of tuberculosis.

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“…In the early period of the AIDS epidemic and before the advent of highly active anti-retroviral therapy (HAART), M. kansasii ranked second behind Mycobacterium avium complex as the most common nontuberculosis mycobacteria (NTM) species isolated from AIDS-associated opportunistic NTM infections (Valainis et al, 1991). Recent studies suggest an increasing incidence of M. kansasii infections in both HIV-positive (Shafer & Sierra, 1992;Witzig et al, 1995) and HIV-negative patients (Bittner et al, 1996). Currently, in many areas of the world, M. kansasii is the most frequent NTM isolated (Bloch et al, 1998;Chobot et al, 1997;Taillard et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Mycobacterium kansasii: antibiotic susceptibility and PCR-restriction analysis of clinical isolates

Telles

Chimara

Ferrazoli

et al. 2005

Journal of Medical Microbiology

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Emergence of Mycobacterium kansasii as the Leading Mycobacterial Pathogen Isolated Over a 20-Year Period at a Midwestern Veterans Affairs Hospital

Cited by 31 publications

References 0 publications

Rapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR

Rapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR

Tuberculin Skin Testing and In Vitro T Cell Responses to ESAT‐6 and Culture Filtrate Protein 10 after Infection withMycobacterium marinumorM. kansasii

Mycobacterium kansasii: antibiotic susceptibility and PCR-restriction analysis of clinical isolates

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