The Combination of Real-Time PCR and HPLC for the Identification of Non-Tuberculous Mycobacteria

Park, Jae Sun; Choi, Jung In; Lim, Jung Hyun; Ahn, Jong Joon; Jegal, Yangjin; Seo, Kwang Won; Won, Seung; Jeon, Jae Bum; Lee, Seon Ho; Kim, Sung Ryul; Jeong, Joseph

doi:10.3343/alm.2013.33.5.349

Cited by 11 publications

(9 citation statements)

References 14 publications

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“…Compared to previous studies of NTM distribution in several tertiary-care hospitals in South Korea, there were a number of differences in distribution of NTM, although the most commonly isolated NTM was generally MAC (Table 3) (8-11, 14, 15, 24, 25). In most studies using molecular methods, M. abscessus was the second-most-common species, but in a high-performance liquid chromatography (HPLC) study, where HPLC is used to identify NTM species by analyzing the mycolic acids in an organism, it ranked only 4th or 5th in prevalence among all of the isolated NTM (15,24). Rather, M. kansasii was the 1st-or 2nd-most-common NTM species in studies using HPLC (15,24).…”

Section: Discussionmentioning

confidence: 99%

“…In most studies using molecular methods, M. abscessus was the second-most-common species, but in a high-performance liquid chromatography (HPLC) study, where HPLC is used to identify NTM species by analyzing the mycolic acids in an organism, it ranked only 4th or 5th in prevalence among all of the isolated NTM (15,24). Rather, M. kansasii was the 1st-or 2nd-most-common NTM species in studies using HPLC (15,24). Considering that the studies using the HPLC method were all performed in the same region, the difference in the epidemiologic data may be representative of the significant geographic variability of NTM.…”

Section: Discussionmentioning

confidence: 99%

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Distribution of Nontuberculous Mycobacteria by Multigene Sequence-Based Typing and Clinical Significance of Isolated Strains

et al. 2014

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bSpecies identification of nontuberculous mycobacteria (NTM) is challenging due to the increasing number of identified NTM species and the lack of standardized testing strategies. The objectives of this study were to investigate the distribution of NTM species recovered from respiratory specimens by multigene sequence-based typing and to evaluate the clinical significance of identified species. Two hundred thirty-two consecutive clinical NTM isolates were subjected to sequencing of multiple genes, including hsp65, rpoB, and 16S-23S rRNA internal transcribed spacer (ITS) sequence. In addition, clinical data from all patients whose specimens had NTM isolates were analyzed to examine clinical virulence and treatment history. Eighteen strains from 227 isolates from 169 patients were successfully identified at the species level by multigene sequence-based typing. Mycobacterium avium complex and M. abscessus complex made up the majority of isolated NTM (88%; 199/227), followed by M. fortuitum complex (4%; 10/227). The pathogenic potential of NTM differs enormously by species, and M. avium complex and M. abscessus complex revealed especially high levels of virulence compared with findings for other NTM species. The results from our work support M. avium complex and M. abscessus complex being the most common NTM species with highly pathogenic potential isolated from clinical respiratory specimens and could be a good resource for molecular epidemiology of NTM species in South Korea.A ll members of the genus Mycobacterium, excepting M. tuberculosis and M. leprae, are considered nontuberculous mycobacteria (NTM) and are frequently isolated from environmental sources, including surface or tap water and soil (1). Currently, more than 160 mycobacterial species/subspecies are listed in the Genus Mycobacterium database (http://www.bacterio.cict.fr/m /mycobacterium.html; accessed on 22 August 2013), and the number of newly identified NTM is increasing. Recent data suggest that the incidence of lung disease caused by NTM with or without predisposing risk factors has been growing worldwide (2-6). However, epidemiologic data on the distribution and clinical significance of NTM lung disease are still scarce because NTM lung disease is not a reportable condition in most countries.In South Korea, NTM was first described as a cause of mycobacterial lung disease in 1981 (7); since then, the frequency of isolation of NTM from clinical specimens has shown a continuous increase. Since the 2000s, the frequency of NTM isolation has been reported to be 12 to 38% of mycobacterial culture-positive specimens, and the number of patients with NTM disease has also been considerable, with a frequency of 8 to 49% of isolated NTM (8-15). The isolation of NTM species from respiratory specimens is not sufficient evidence for the diagnosis of clinically significant NTM lung disease, since there are also clinical, radiographic, and microbiological criteria (1).Recently, various genotype-based methods for the identification of NTM have been developed to ra...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Distribution of Nontuberculous Mycobacteria by Multigene Sequence-Based Typing and Clinical Significance of Isolated Strains

et al. 2014

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“…The real-time PCR assays are advantageous because of its rapidity and high sensitivity. Furthermore, the specificity of the real-time PCR can be enhanced by combination with HPLC, which is a useful tool to discriminate NTM at the species level, although it requires specific equipment and technical expertise [113]. Furthermore, multiplex real-time PCR assay combined with melting curve analysis is also an accurate, rapid and effective tool for the mycobacterial identification from cultures [114].…”

Section: Molecular-based Identification Methodsmentioning

confidence: 99%

Overview of Non Tuberculosis Mycobacterial Lung Diseases

Diseases¹

2018

Mycobacterium - Research and Development

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Nontuberculosis mycobacteria (NTM) are ubiquitous in nature and opportunistically infect different animals, including humans. Currently, NTM is emerging as an important cause of pulmonary infection among both immunocompromised and immunocompetent persons worldwide. The clinical relevance of pulmonary NTM varies among species while showing geographical heterogeneity in distribution as well as pathogenicity. The outcome of the respiratory NTM disease is a consequence of a complex interplay between microbial factors and host susceptibility. Furthermore, HIV infection, cystic fibrosis, cancer, underlying chronic lung disease and history of tuberculosis (TB) may be associated as risk factors for active nontuberculosis pulmonary diseases (NTMPD). The diagnosis of NTMPD requires the presence of symptoms, radiographic evidences, microscopic observations and definitive laboratory diagnostics. Lung infections resulted from a clinically significant NTM species should be treated with appropriate antimicrobial regimen. M. leprae and M. lepraemurium are uncultivable and require the intracellular milieu for survival and propagation [4]. The obligatory causative agents of the genus Mycobacterium, responsible for TB are classified into Mycobacterium tuberculosis complex (MTC). It comprises M. tuberculosis, M.bovis, M. africanum, M. microti [5], M. canettii [6], M. caprae [7], M. pinnipedii [8], M. mungi [9], M. orygis [10] and M. suricattae [11] species. M. tuberculosis, M. africanum, M. canettii and M. orygis cause TB primarily in human [4, 10], whereas M. bovis [12], M. microti [13], M. caprae [7] M. pinnipedii [8], M. mungi [9] and M. suricattae [11] infect cattle, domestic animals, goats, seals, mongooses and meerkats, respectively, and animal tuberculosis can also be zoonotic [14, 15]. However, geographical variation of the MTC species distribution has been identified. As an example, M. africanum is a common cause of human pulmonary TB (39%) as much as M. tuberculosis (55%) in West Africa [16]. In Ghana, 3% of pulmonary TB cases are represented by M. bovis, while 20% are M. africanum and 73% are M. tuberculosis [17].Other medically important mycobacteria such as M. avium, M. intracellulare complex, M. kansasii, M. marinum, M. fortuitum, M. chelonae complex, M. abscessus and M. scrofulaceum are known as nontuberculosis mycobacteria (NTM) species or atypical mycobacteria or mycobacteria other than tuberculosis (MOTT). They are responsible for diseases including lymphadenitis in children, chronic pulmonary diseases, skin and soft-tissue diseases and infections of the skeletal system [18]. NTM are ubiquitous in nature and are widely distributed in water, soil and animals. Among prevailing NTM species, only a few species have a clinical impact on humans as opportunistic pathogens [19]. M. avium complex (MAC), M. abscessus, M. kansasii, M. fortuitum, M. chelonae, M. szulgai, M. triviale and M. scrofulaceum are common NTM species that cause pulmonary diseases in human [20]. Additionally, M. riyadhense was recently proposed as a cau...

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“…However, these methods have a requirement for the presence of mycobacterial cells 10 7 ‐10 8 to be effective that makes this process necessitates the 14‐21 culture day. HPLC is quick and repeatable method based on chromatographic method to determine mycolic acid esters present in the cell wall of mycobacteria . There were a limited number of study for HPLC analysis for mycobacterium species.…”

Section: Introductionmentioning

confidence: 99%

Determination of antibiotic resistance and high‐performance liquid chromatography profiles for Mycobacterium species

Özer

Yula²,

Doğan

et al. 2018

Clinical Laboratory Analysis

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The Combination of Real-Time PCR and HPLC for the Identification of Non-Tuberculous Mycobacteria

Cited by 11 publications

References 14 publications

Distribution of Nontuberculous Mycobacteria by Multigene Sequence-Based Typing and Clinical Significance of Isolated Strains

Distribution of Nontuberculous Mycobacteria by Multigene Sequence-Based Typing and Clinical Significance of Isolated Strains

Overview of Non Tuberculosis Mycobacterial Lung Diseases

Determination of antibiotic resistance and high‐performance liquid chromatography profiles for Mycobacterium species

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