Detection of non-tuberculous mycobacteria in hospital water by culture and molecular methods

Hussein, Ziyad; Landt, Olfert; Wirths, Beate; Wellinghausen, Nele

doi:10.1016/j.ijmm.2008.07.004

Cited by 50 publications

(56 citation statements)

References 28 publications

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“…Multiplexing strategies allow for further improvement of diagnostics in terms of rapidity and efficiency, and multiplex real-time PCR represents a reliable alternative. Among the various multiplex PCR assays reported thus far (20)(21)(22)(23)(24)(25)(26), many are conventional PCRs or use melting curve analysis and need additional interpretation; some have not been used directly on clinical samples, and others seem to have some specificity or sensitivity issues. A recently published duplex real-time PCR assay for the detection of M. tuberculosis and M. avium complexes on human respiratory specimens has been shown to be reliable and cost-effective (27), but it could have an added value by including an additional target for genus detection.…”

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Detection of Mycobacteria, Mycobacterium avium Subspecies, and Mycobacterium tuberculosis Complex by a Novel Tetraplex Real-Time PCR Assay

et al. 2015

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bMycobacterium tuberculosis complex, Mycobacterium avium, and many other nontuberculous mycobacteria are worldwide distributed microorganisms of major medical and veterinary importance. Considering the growing epidemiologic significance of wildlife-livestock-human interrelation, developing rapid detection tools of high specificity and sensitivity is vital to assess their presence and accelerate the process of diagnosing mycobacteriosis. Here we describe the development and evaluation of a novel tetraplex real-time PCR for simultaneous detection of Mycobacterium genus, M. avium subspecies, and M. tuberculosis complex in an internally monitored single assay. The method was evaluated using DNA from mycobacterial (n ‫؍‬ 38) and nonmycobacterial (n ‫؍‬ 28) strains, tissues spiked with different CFU amounts of three mycobacterial species (n ‫؍‬ 57), archival clinical samples (n ‫؍‬ 233), and strains isolated from various hosts (n ‫؍‬ 147). The minimum detectable DNA amount per reaction was 50 fg for M. bovis BCG and M. kansasii and 5 fg for M. avium subsp. hominissuis. When spiked samples were analyzed, the method consistently detected as few as 100 to 1,000 mycobacterial CFU per gram. The sensitivity and specificity values for the panel of clinical samples were 97.5 and 100% using a verified culture-based method as the reference method. The assays performed on clinical isolates confirmed these results. This PCR was able to identify M. avium and M. tuberculosis complex in the same sample in one reaction. In conclusion, the tetraplex real-time PCR we designed represents a highly specific and sensitive tool for the detection and identification of mycobacteria in routine laboratory diagnosis with potential additional uses. T he genusMycobacterium encompasses a large number of worldwide distributed pathogenic and opportunistic bacteria responsible for a broad range of infectious diseases. Mycobacterium tuberculosis complex, Mycobacterium avium, and many other nontuberculous mycobacteria (NTM) are of major medical and veterinary significance. Human tuberculosis (TB) due to infection with members of M. tuberculosis complex, especially with M. tuberculosis, remains an important health problem according to annual reports of the World Health Organization (WHO). Animal or bovine TB (bTB), caused primarily by Mycobacterium bovis but also by Mycobacterium caprae, poses serious socioeconomic and health threats (1) and is a notifiable disease listed under the World Organization for Animal Health (OIE) Terrestrial Animal Health Code (TAHC). Although bTB is controlled in most developed countries, its eradication has not been fully accomplished due to a lack of sensitivity of diagnostic tools and to the persistence of infection in extensive farming and wild populations (2). The disease remains a significant problem in many countries with less developed laboratory capacity (1). Accordingly, human TB caused by M. bovis or M. caprae is supposed to be infrequent in developed regions but is relatively common and possibly underestimated in ...

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Detection of Mycobacteria, Mycobacterium avium Subspecies, and Mycobacterium tuberculosis Complex by a Novel Tetraplex Real-Time PCR Assay

et al. 2015

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“…Mycobacterial infections are believed to be a common cause of SGI; however, as noted in our findings, staining for these organisms is often negative and although tissue culture increases the chances of identifying a specific infectious agent, it is time-consuming. Bartralot et al have stressed that infections caused by different species of NTM cannot be differentiated on histopathologic grounds (1).Because of its high sensitivity and specificity, the PCR is now widely employed to amplify NTM-specific nucleic acids in a variety of clinical and environmental specimens (4,6,7,11). The suspension array system has been used for the characterization and assessment of mycobacterial infections, as previously reported (12-14).…”

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confidence: 99%

“…Clinical presentations include infections of the skin and soft tissue, lymph nodes, joints, and lungs. Disseminated infections may arise following the contamination of traumatic or surgical wounds with contaminated water or other materials (5,6). Most species of NTM have been associated with skin and soft tissue infections, but rapidly growing mycobacteria (Mycobacterium fortuitum, M. chelonae, and M. abscessus), M. marinum, and M. ulcerans have been the ones most frequently described.…”

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confidence: 99%

“…Some of these organisms exhibit geographic restrictions. The timely and accurate diagnosis of cutaneous NTM infections is important in that different species of these organisms have different antibiotic susceptibilities (4,6,7). Current laboratory techniques for identifying mycobacteria to the species level include growth in broth, nucleic acid probing, and sequencing, as well as subculture on solid medium for phenotypic testing, each of which is time intensive (4,7,8).…”

mentioning

confidence: 99%

“…Because of its high sensitivity and specificity, the PCR is now widely employed to amplify NTM-specific nucleic acids in a variety of clinical and environmental specimens (4,6,7,11). The suspension array system has been used for the characterization and assessment of mycobacterial infections, as previously reported (12)(13)(14).…”

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Mycobacterium smegmatis in Skin Biopsy Specimens from Patients with Suppurative Granulomatous Inflammation

Lü

Xia

et al. 2013

J Clin Microbiol

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Formalin-fixed, paraffin-embedded skin biopsy specimens, including 72 suppurative granulomatous inflammation (SGI) and 47 non-SGI controls, were tested for mycobacteria by using a broad-range PCR and a suspension array identification system. Mycobacterium smegmatis was detected in 13 (18.1%) of the SGI skin biopsy specimens, which was significantly more than 2 (4.3%) in the controls (odds ratio, 5.73; 95% confidence interval, 1.21 to 27.06; P ‫؍‬ 0.028).G ranulomatous inflammation of the skin may be broadly classified as infectious or noninfectious, with the presence of a suppurative inflammatory component often associated with the former (1-3). Suppurative granulomatous inflammation (SGI) has been defined as inflammation within the dermis and subcutis consisting of neutrophils, various numbers of eosinophils, and the presence of a histiocytic population of cells (1, 2). Multinucleated giant cells are usually present without obvious foreign material. A diagnosis may be made histologically; however, prior antibiotic exposure may "sterilize" the tissue, leading to a delayed diagnosis and increased morbidity. Tissue culture is the diagnostic gold standard, but when a mycobacterial infection is responsible, cultures often take weeks to grow (4, 5).To date, about one-third of the species of nontuberculous mycobacteria (NTM) described have been associated with human disease (5). Clinical presentations include infections of the skin and soft tissue, lymph nodes, joints, and lungs. Disseminated infections may arise following the contamination of traumatic or surgical wounds with contaminated water or other materials (5, 6). Most species of NTM have been associated with skin and soft tissue infections, but rapidly growing mycobacteria (Mycobacterium fortuitum, M. chelonae, and M. abscessus), M. marinum, and M. ulcerans have been the ones most frequently described. Some of these organisms exhibit geographic restrictions. The timely and accurate diagnosis of cutaneous NTM infections is important in that different species of these organisms have different antibiotic susceptibilities (4, 6, 7). Current laboratory techniques for identifying mycobacteria to the species level include growth in broth, nucleic acid probing, and sequencing, as well as subculture on solid medium for phenotypic testing, each of which is time intensive (4,7,8).We have developed a MycoID assay that incorporates broadrange PCR amplification with suspension array detection to identify 17 mycobacterial complexes, groups, and species in a single reaction with a detection limit of 50 to 500 copies/g of tissue (8). In this study, we have employed the MycoID procedure to detect and identify mycobacterial species in formalin-fixed, paraffin-embedded (FFPE) skin biopsy specimens exhibiting SGI and non-SGI controls.(This study was presented in part at the Association for Molecular Pathology Annual Meeting, Grapevine, TX, 17 to 19 November 2011.)Study materials. The study FFPE tissue specimens consisted of 119 skin biopsy specimens obtained from Beijing Children...

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Infektionen mit nichttuberkulösen Mykobakterien bei Patienten mit Immundefekten

Rupp

Schaaf

2011

Pneumologe

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Leitthema Der Nachweis von nichttuberkulösen Mykobakterien (NTM) über-steigt in vielen industrialisierten Län-dern die Nachweisraten von M. tuberculosis (MTB). So wurden in einerPopulationsstudie bei Patienten mit pulmonalen Infiltraten NTM deutlich häufiger nachgewiesen als MTB (7,2/100.000 vs. 2,8/100.000 Patienten) [1]. Die Zahl der NTM-Neuerkrankungen ist seit vielen Jahren mit 1-2/100.000 Personen relativ konstant [2].

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Detection of non-tuberculous mycobacteria in hospital water by culture and molecular methods

Cited by 50 publications

References 28 publications

Detection of Mycobacteria, Mycobacterium avium Subspecies, and Mycobacterium tuberculosis Complex by a Novel Tetraplex Real-Time PCR Assay

Detection of Mycobacteria, Mycobacterium avium Subspecies, and Mycobacterium tuberculosis Complex by a Novel Tetraplex Real-Time PCR Assay

Mycobacterium smegmatis in Skin Biopsy Specimens from Patients with Suppurative Granulomatous Inflammation

Infektionen mit nichttuberkulösen Mykobakterien bei Patienten mit Immundefekten

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