Simultaneous detection of Mycobacterium tuberculosis complex and nontuberculous mycobacteria in respiratory specimens

Hwang, Sang Mee; Lim, Mi Suk; Hong, Yun Ji; Kim, Taek Soo; Park, Kyoung Un; Song, Jae-Joon; Lee, Jae Ho; Kim, Eui Chong

doi:10.1016/j.tube.2013.07.007

Cited by 29 publications

(27 citation statements)

References 19 publications

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“…marinum in the skin and soft tissue category, although the absolute numbers were small (Table 3). Our finding of the simultaneous occurrence of MTC and NTM in 17% of adequately analyzed specimens accords with those of Hwang et al [9], who used various direct molecular techniques plus culturing of respiratory specimens.…”

Section: Discussionsupporting

confidence: 92%

Two-Tube Multiplex PCR for Genotyping Tuberculous and Nontuberculous Mycobacterial Species in Pathology Specimens

Dictor¹,

Warenholt²,

Łukasiewicz³

2018

AiM

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Background: The incidence of mycobacterial infection, in particular M. tuberculosis complex (MTC), is increasing in some Western countries, while nontuberculous mycobacteria (NTM) may be recognized more frequently in clinical specimens worldwide. The clinical scenario and available histopathology alone are often insufficient to separate these two categories of mycobacterial disease, whose behavior and treatment differ. In particular, NTM may be clinically unsuspected in pathological specimens and the opportunity for culturing missed. Methods: We developed two multiplex PCR assays, which distinguish MTC from NTM by detecting the IS6110 insert in the first tube and discriminating up to 14 NTM reference strains in the second by targeting the 16S-23S rRNA internal transcribed spacer. Test material included 594 routine clinical specimens with diverse pathology; many were granulomas unrelated to mycobacterial infection. About 75% were formalin-fixed paraffin blocks, the remainder mainly cytologic imprints or aspirates on FTA cards submitted on suspicion of mycobacterial infection either to avoid frozen sectioning (with the attendant risk of aerosolisation) or at the time of fine needle aspiration. Results: The paraffinized material yielded 53 MTC positives and the cytological 21 positives. A subset consisting of 337 specimens was also analyzed for NTM and yielded 51 positives. The frequency of simultaneous NTM infection in tuberculous patients was about 17%. Mycobacterium avium complex represented the dominant NTM species overall, showed a predilection for lung and lymph node, and together with M. haemophilum were the second most frequent NTM just behind M. ulcerans/M. marinum in skin and soft tissue, the category displaying the largest NTM diversity. Conclusions: Cytological and deparaffinized tissue analyzed in a new two-tube multiplex PCR allows for specific discrimination of causative agents in mycobacterial infection. MTC is readily distinguished from NTM for appropriate therapy, and NTM presumptively diagnosed at the species level allows appropriate choices of antimicrobials.

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Section: Discussionsupporting

confidence: 92%

Two-Tube Multiplex PCR for Genotyping Tuberculous and Nontuberculous Mycobacterial Species in Pathology Specimens

Dictor¹,

Warenholt²,

Łukasiewicz³

2018

AiM

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“…Many regions of the mycobacterial genome have been used in molecular diagnosis of TB, such as IS6110, 16S rRNA, hsp65, rpoB, sdaA, devR and mpt64 (Manjunath et al, 1991;Lachnik et al, 2002;Kim et al, 2004;Chakravorty et al, 2005;Negi et al, 2007;Hwang et al, 2013;Nimesh et al, 2013). mpt64 is found as a single copy in the genome of MTBC species (Lee et al, 1994), and has been used for both pulmonary and extrapulmonary TB diagnosis (Tan et al, 1995;Dar et al, 1998;Martins et al, 2000;Bhanu et al, 2005;Therese et al, 2005;Takahashi & Nakayama, 2006;Dil-Afroze et al, 2008;Kusum et al, 2012;Sethi et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of Mycobacterium tuberculosis complex by real-time PCR in sputum samples and its use in the routine diagnosis in a reference laboratory

Pinhata

Cergole-Novella²,

Carmo³

et al. 2015

Journal of Medical Microbiology

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Tuberculosis (TB) is an infectious disease of global distribution, constituting a serious public health problem in Brazil. Sã o Paulo State, located in the south-east of Brazil, notified 16 580 new TB cases in 2013. The Instituto Adolfo Lutz is a public health reference laboratory for TB diagnosis for all the State. Considering that rapid and accurate diagnosis is essential for TB control, the aim of this study was to evaluate the use of an in-house real-time (RT)-PCR assay targeting the mpt64 gene in the routine diagnosis of TB, and to compare this technique with smear microscopy and culture. From August 2012 to October 2013, 715 sputum samples from 657 patients were included in the study. Smear microscopy, culture, phenotypic and PRAhsp65 identification of mycobacteria, and mpt64 RT-PCR were performed. With respect to confirmed TB cases (n562/657; 9.4 %), smear microscopy had a sensitivity of 82.3 %. Culture and RT-PCR showed the same sensitivity, i.e. 90.3 %. Specificity was 99.7, 99.4 and 98.6 % for smear microscopy, culture and RT-PCR, respectively. mpt64 RT-PCR showed high sensitivity and specificity for the detection of Mycobacterium tuberculosis complex in sputum samples. This technique can be deployed in laboratories that do not have a rapid test for TB available, enabling the performance of TB diagnosis in up to 5 h.

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“…Th e dual recovery of M tuberculosis and NTM has also been noted. [5][6][7][8][9][10] Th e underlying bronchial conditions or milieu promoting mycobacterial acquisition and persistence in the lung are likely very similar, if not identical, regardless of the primary mycobacterial pathogen. Th e challenge is to determine the signifi cance of an NTM isolated in this context, as identifi cation of clinically relevant NTM isolates requires satisfying a combination of clinical, radiologic, and microbiologic criteria, which is neither facile nor inconsequential, as it could obligate the patient to further prolonged antimycobacterial therapy.…”

Section: Discussionmentioning

confidence: 99%

“…[5][6][7][8][9][10] To date, however, there has not been suffi cient data amassed to make confi dent conclusions about optimal management of patients with nontuberculous mycobacterial (NTM) isolation during a course of TB therapy. Th e major shortcomings of the available studies have been either insuffi cient duration of patient follow-up or inadequate, individual patient clinical assessment.…”

mentioning

confidence: 99%

“…Th e major shortcomings of the available studies have been either insuffi cient duration of patient follow-up or inadequate, individual patient clinical assessment. [5][6][7][8][9][10] We recently reported microbiologic treatment outcomes for 180 patients with MAC lung disease. 11 A substantial number of those patients had concomitant isolation of other NTM with MAC, primarily MAA.…”

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confidence: 99%

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The Significance of Mycobacterium abscessus Subspecies abscessus Isolation During Mycobacterium avium Complex Lung Disease Therapy

Griffith

Philley

Benwill

et al. 2015

Chest

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Simultaneous detection of Mycobacterium tuberculosis complex and nontuberculous mycobacteria in respiratory specimens

Cited by 29 publications

References 19 publications

Two-Tube Multiplex PCR for Genotyping Tuberculous and Nontuberculous Mycobacterial Species in Pathology Specimens

Two-Tube Multiplex PCR for Genotyping Tuberculous and Nontuberculous Mycobacterial Species in Pathology Specimens

Rapid detection of Mycobacterium tuberculosis complex by real-time PCR in sputum samples and its use in the routine diagnosis in a reference laboratory

The Significance of Mycobacterium abscessus Subspecies abscessus Isolation During Mycobacterium avium Complex Lung Disease Therapy

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