Rapid and Accurate Identification of Mycobacterium tuberculosis Complex and Common Non-Tuberculous Mycobacteria by Multiplex Real-Time PCR Targeting Different Housekeeping Genes

Esfahani, Bahram Nasr; Yazdi, Hadi Rezaei; Moghim, Sharareh; Safaei, Hajieh Ghasemian; Zarkesh-Esfahani, Hamid

doi:10.1007/s00284-012-0188-2

Cited by 24 publications

(17 citation statements)

References 27 publications

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“…M. mucogenicum, M. senegalense, M. genavense, and M. simiae are also opportunistic pathogens that are capable of causing disease in those with impaired immune systems [20] . M. mucogenicum was also detected in a similar study by Nasr Esfahani et al [21] in central Iran, where the dominant isolates were M. fortuitum, M. chelonae-like organism, and M. mucogenicum. A similar study by Mohajeri et al [22] evaluating the frequency of NTM in drinking water supplies revealed NTM in 32% of 110 samples, with M. gastri as the most common species.…”

Section: Discussionsupporting

confidence: 68%

Isolation and identification of Nontuberculous mycobacteria from environmental water samples in the northeast of Iran

Akbari¹,

Zare²,

Ghazvini³

2019

mjima

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Giriş: Çevresel insan enfeksiyonu kaynağı olarak son zamanlarda giderek artan düzeyde dikkat çeken ve toplamda 150'den fazla türü olan nontüberküloz mikobakterilere (NTM) verilen önem artmaktadır. Bu çalışmanın amacı, İran'ın kuzeydoğusunda çevresel NTM türlerini izole etmek ve tanımlamaktı. Gereç ve Yöntem: Razavi Horasan eyaletinin 73 şehrindeki içme suyu dağıtım ağları ve su kaynaklarından toplanan 344 su örneği kültür yoluyla incelendi. Nontüberküloz mikobakteriler tanımlaması ayrıca hsp65 geninin polimeraz zincir reaksiyonu amplifikasyonu, sekanslama ve veri analizi yoluyla da yapıldı. İstatistiksel analiz, SPSS 16.0 kullanılarak Pearson korelasyon katsayısı testi ve iki örnek t-testi ile yapıldı. istatistiksel anlamlılık seviyesi 0,05 olarak belirlendi. Bulgular: Nontüberküloz mikobakteriler, kültürde su örneklerinin %10,46'sında (36/244) tespit edilmiştir. En yaygın NTM türü Mycobacterium gordonae (13/36) olarak belirlenmiştir. Diğer saptanan türler M. mucogenicum, M. senegalense, M. gadium, M. genavense, M. simiae, M. frederiksbergense, M. flouranthenivorans, M. neoaurum, ve M. pallens'di. Ayrıca, suda NTM varlığı ile nitrat seviyesi, türbidite, pH ve su kaynağının Introduction: The importance of nontuberculous mycobacteria (NTM), a group including over 150 species, has recently received increased attention as an environmental source of human infection. This study aimed to isolate and identify NTM species in the environment in the northeast of Iran. Materials and Methods: A total of 344 water samples were obtained from water resources and drinking water distribution networks in 73 cities in the Khorasan Razavi province and were examined using culture. Nontuberculous mycobacteria identification was also accomplished by polymerase chain reaction amplification of the hsp65 gene, sequencing and data analysis. Statistical analysis was performed by Pearson's correlation coefficient test and two-sample T-test using SPSS 16.0. Statistical significance was determined at a significance level of 0.05. Results: Nontuberculous mycobacteria were identified in 10.46% of the water samples (36/344) by culture. The most frequent NTM species was Mycobacterium gordonae (13/36). Other Mycobacterium species detected included M . mucogenicum, M. senegalense, M. gadium, M. genavense, M. simiae, M. frederiksbergense, M. flouranthenivorans, M. neoaurum, and M. pallens. Statistically significant correlations were observed between the presence of NTM in water and nitrate level, turbidity, pH, and age of water resource (p<0.001). The relation between NTM presence and depth of the water resource was not statistically significant (p>0.05). Conclusion: It was concluded that NTM water contamination was not very high in this area, but further studies are needed to investigate the sources of contamination and origins of these species.

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Section: Discussionsupporting

confidence: 68%

Isolation and identification of Nontuberculous mycobacteria from environmental water samples in the northeast of Iran

Akbari¹,

Zare²,

Ghazvini³

2019

mjima

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“…3,5,7e9,13e34 The turnaround time for commercial MPT64-based ICTs is 0.5e1 h, whereas it might be several days for molecular methods and several weeks for biochemical method. 35 In Figure 1 Study selection process. MPT-64 based tests for identification of MTC addition to rapidity, commercial MPT64-based ICTs are less expensive and technologically simper than molecular methods and biochemical method.…”

Section: Discussionmentioning

confidence: 98%

Commercial MPT64-based tests for rapid identification of Mycobacterium tuberculosis complex: A meta-analysis

Yin¹,

Zheng

et al. 2013

Journal of Infection

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“…Previously reported PCR methods for genus detection seem to have different drawbacks apparently avoided by this new PCR. As alignment between published primers/probes and genomic targets used by other researchers (16S rRNA, ITS, and heat shock protein 65) suggested, some might not detect DNA from various mycobacteria (37,38), others would detect nonmycobacterial DNA (20,21,23,25,32), and others might combine both sensitivity and specificity limitations (22,24,26,39). For M. avium subspecies detection, we used the multicopy element IS1311, a target previously used in multiplex real-time PCR assays (25) that, as far as we know, is M. avium subspecies specific (5).…”

Section: Discussionmentioning

confidence: 99%

“…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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confidence: 99%

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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Rapid and Accurate Identification of Mycobacterium tuberculosis Complex and Common Non-Tuberculous Mycobacteria by Multiplex Real-Time PCR Targeting Different Housekeeping Genes

Cited by 24 publications

References 27 publications

Isolation and identification of Nontuberculous mycobacteria from environmental water samples in the northeast of Iran

Isolation and identification of Nontuberculous mycobacteria from environmental water samples in the northeast of Iran

Commercial MPT64-based tests for rapid identification of Mycobacterium tuberculosis complex: A meta-analysis

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

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