Use of the GenoType Mycobacterium CM and AS Assays To Analyze 76 Nontuberculous Mycobacterial Isolates from Greece

Gitti, Zoe; Neonakis, Ioannis K.; Fanti, Garyfallia; Kontos, Fanourios; Maraki, Sofia; Tselentis, Yiannis

doi:10.1128/jcm.02088-05

Cited by 39 publications

(24 citation statements)

References 9 publications

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“…Among the three probes used in the AccuProbe technique (MAC, M. avium, and M. intracellulare), a lack of specificity of the MAC and M. intracellulare probes has been reported. In addition, misidentifications and no identification have been reported with the DNA strip technologies (9,19,25,33). The MALDI-TOF MS strategy can differentiate between the two species of the MAC in one step, unlike the AccuProbe technique, which needs two probes to precisely identify these bacterial species.…”

Section: Discussionmentioning

confidence: 99%

Rapid Identification of Mycobacterial Whole Cells in Solid and Liquid Culture Media by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

et al. 2010

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Mycobacterial identification is based on several methods: conventional biochemical tests that require several weeks for accurate identification, and molecular tools that are now routinely used. However, these techniques are expensive and time-consuming. In this study, an alternative method was developed using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). This approach allows a characteristic mass spectral fingerprint to be obtained from whole inactivated mycobacterial cells. We engineered a strategy based on specific profiles in order to identify the most clinically relevant species of mycobacteria. To validate the mycobacterial database, a total of 311 strains belonging to 31 distinct species and 4 species complexes grown in Löwenstein-Jensen (LJ) and liquid (mycobacterium growth indicator tube [MGIT]) media were analyzed. No extraction step was required. Correct identifications were obtained for 97% of strains from LJ and 77% from MGIT media. No misidentification was noted. Our results, based on a very simple protocol, suggest that this system may represent a serious alternative for clinical laboratories to identify mycobacterial species.

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Section: Discussionmentioning

confidence: 99%

Rapid Identification of Mycobacterial Whole Cells in Solid and Liquid Culture Media by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

et al. 2010

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“…Genomic DNA from heat-killed AFB isolates was extracted. The GenoType Mycobacterium CM/AS assay (Hain Lifesciences) was performed following the manufacturer's protocol (45). The reaction mixture consisted of 2 l of MgCl 2 , 5 l of 10ϫ buffer, 35 l of biotinylated primer-nucleotide mixture, 0.2 l of HotStar Taq (Qiagen, Hilden, Germany), 2.8 l of nuclease-free water, and 5 l of DNA, in a total volume of 50 l. PCR was performed as follows: 95°C for 15 min, 10 cycles of 95°C for 30 s and 58°C for 2 min, 20 cycles of 95°C for 25 s and 53°C for 40 s, and 20 cycles of 70°C for 8 min.…”

Section: Methodsmentioning

confidence: 99%

Isolation of Nontuberculous Mycobacteria from the Environment of Ghanian Communities Where Buruli Ulcer Is Endemic

Aboagye

Ampah

Nakobu

et al. 2016

Appl Environ Microbiol

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This study aimed to isolate nontuberculous mycobacterial species from environmental samples obtained from some selected communities in Ghana. To optimize decontamination, spiked environmental samples were used to evaluate four decontamination solutions and supplemented media, after which the best decontamination solution and media were used for the actual analysis. The isolates obtained were identified on the basis of specific genetic sequences, including heat shock protein 65, IS2404, IS2606, rpoB, and the ketoreductase gene, as needed. Among the methods evaluated, decontamination with 1 M NaOH followed by 5% oxalic acid gave the highest rate of recovery of mycobacteria (50.0%) and the lowest rate of contamination (15.6%). The cultivation medium that supported the highest rate of recovery of mycobacteria was polymyxin B-amphotericin B-nalidixic acidtrimethoprim-azlocillin-supplemented medium (34.4%), followed by isoniazid-supplemented medium (28.1%). Among the 139 samples cultivated in the main analysis, 58 (41.7%) yielded mycobacterial growth, 70 (50.4%) had no growth, and 11 (7.9%) had all inoculated tubes contaminated. A total of 25 different mycobacterial species were identified. Fifteen species (60%) were slowly growing (e.g., Mycobacterium ulcerans, Mycobacterium avium, Mycobacterium mantenii, and Mycobacterium malmoense), and 10 (40%) were rapidly growing (e.g., Mycobacterium chelonae, Mycobacterium fortuitum, and Mycobacterium abscessus). The occurrence of mycobacterial species in the various environmental samples analyzed was as follows: soil, 16 species (43.2%); vegetation, 14 species (38.0%); water, 3 species (8.0%); moss, 2 species (5.4%); snail, 1 species (2.7%); fungi, 1 species (2.7%). This study is the first to report on the isolation of M. ulcerans and other medically relevant nontuberculous mycobacteria from different environmental sources in Ghana. IMPORTANCEDiseases caused by mycobacterial species other than those that cause tuberculosis and leprosy are increasing. Control is difficult because the current understanding of how the organisms are spread and where they live in the environment is limited, although this information is needed to design preventive measures. Growing these organisms from the environment is also difficult, because the culture medium becomes overgrown with other bacteria that also live in the environment, such as in soil and water. We aimed to improve the methods for growing these organisms from environmental sources, such as soil and water samples, for better understanding of important mycobacterial ecology.

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“…[1] The introduction of molecular techniques has facilitated the detection and identiÞ cation of new NTM species, the role of which is under constant evaluation. [2] Mycobacterium malmoense is a slowly growing NTM that has attracted global interest due to its increased pathogenicity. [3] It is most frequently isolated in cool climates such as Northern and Central Europe.…”

Section: Abstract: Greece Mycobacterium Malmoense Nontuberculous Mmentioning

confidence: 99%

“…Others include Absidia, Mucor, Apophyomyces and Saksenaea. [1,2] Renal involvement in disseminated mucormycosis occurs in up to 19% of patients. [3] Isolated renal zygomycosis (mucormycosis) is extremely rare.…”

Section: Abstract: Apomycophyses Elegans Renal Mucormycosismentioning

confidence: 99%

Isolation of <i> Mycobacterium malmoense</i> in the island of Crete, Greece

Kourbeti¹,

Neonakis²,

Gitti³

et al. 2008

Indian J Med Microbiol

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Use of the GenoType Mycobacterium CM and AS Assays To Analyze 76 Nontuberculous Mycobacterial Isolates from Greece

Cited by 39 publications

References 9 publications

Rapid Identification of Mycobacterial Whole Cells in Solid and Liquid Culture Media by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Rapid Identification of Mycobacterial Whole Cells in Solid and Liquid Culture Media by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Isolation of Nontuberculous Mycobacteria from the Environment of Ghanian Communities Where Buruli Ulcer Is Endemic

Isolation of <i> Mycobacterium malmoense</i> in the island of Crete, Greece

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