Rapid differentiation of mycobacteria by simplex real-time PCR with melting temperature calling analysis

Yin, Xiaomao; Wang, Q.

doi:10.1111/jam.12884

Cited by 2 publications

(2 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although semi-automated culture systems have reduced the time required for the tests, traditional tests for identification between MTB and NTM species still suffer from disadvantages (Kazumi and Mitarai, 2012;Şamlı and İlki, 2016;Gholoobi et al, 2014). The development of molecular diagnostic techniques has significantly increased sensitivity and reduced diagnosis time (Jung et al, 2016;Lin et al, 2015). PCR-based assays have been used to detect mycobacterial DNA with high sensitivity and specificity (Mackay, 2004;Kwon and Koh, 2014).…”

Section: Discussionmentioning

confidence: 99%

Comparison of the Three Molecular Diagnostic Assays for Molecular Identification ofMycobacterium tuberculosisand Nontuberculous Mycobacteria Species in Sputum Samples

Bae

Park

Kim

et al. 2020

BSL

View full text Add to dashboard Cite

Mycobacterium tuberculosis (MTB) continues to be one of the main causative agents of tuberculosis (TB); moreover, the incidence of nontuberculous mycobacteria (NTM) infections has been rising gradually in both immunocompromised and immunocompetent patients. Precise and rapid detection and identification of MTB and NTM in respiratory specimens are thus important for MTB infection control. Molecular diagnostic methods based on the nucleic acid amplification test (NAAT) are known to be rapid, sensitive, and specific compared to the conventional acid-fast bacilli (AFB) smear and mycobacterial culture methods. In the present study, the clinical performances of three commercial molecular diagnostic assays, namely TB/NTM PCR (Biocore), MolecuTech Real MTB-ID ® (YD Diagnostics), and REBA Myco-ID ® (YD Diagnostics), were evaluated with a total of 92 respiratory specimens (22 AFB smear positives and 67 AFB smear negatives). The sensitivity and specificity of TB/NTM PCR were 100% and 75.81%, respectively. The corresponding values of MolecuTech Real MTB-ID ® and REBA Myco-ID® were 56.52% and 90.32%, and 56.52% and 82.26%, respectively. TB/NTM PCR showed the highest sensitivity; however, the concordant rate was 10% compared with sequence analysis. Although MolecuTech Real MTB-ID ® showed lower sensitivity, its specificity was the highest among the three methods. REBA Myco-ID ® allowed accurate classification of NTM species; therefore, it was the most specific diagnostic method. Of the three PCR-based methods, MolecuTech Real MTB-ID ® showed the best performance. This method is expected to enable rapid and accurate identification of MTB and NTM.

show abstract

Section: Discussionmentioning

confidence: 99%

Comparison of the Three Molecular Diagnostic Assays for Molecular Identification ofMycobacterium tuberculosisand Nontuberculous Mycobacteria Species in Sputum Samples

Bae

Park

Kim

et al. 2020

BSL

View full text Add to dashboard Cite

show abstract

“…Our mPCR data based on a large number of MGIT cultures of Mycobacterium species showed that this assay targeting the oxyR-ahpC intergenic region and the variable region of rpoB gene [18] is a rapid, cost-effective and efficient way to differentiate MTB from NTM, and is also capable of identifying mixed MTB + NTM cultures. Other investigators have used more expensive real-time PCR formats with/without expensive probe primers [21, 22]. Although matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has also been used for identification and differentiation of various mycobacteria, the method requires growth on time-consuming solid media for more accurate identification [23, 24].…”

Section: Discussionmentioning

confidence: 99%

Diversity of Nontuberculous Mycobacteria in Kuwait: Rapid Identification and Differentiation of <b><i>Mycobacterium</i></b> Species by Multiplex PCR, INNO-LiPA Mycobacteria v2 Assay and PCR Sequencing of rDNA

Ahmad¹,

Mokaddas²

2019

Med Princ Pract

View full text Add to dashboard Cite

Objective: Nontuberculous mycobacteria (NTM) often cause disease that is clinically indistinguishable from tuberculosis. Specific identification is important as treatment varies according to Mycobacterium species causing the infection. This study used multiplex PCR (mPCR) assay for rapid differentiation of mycobacterial growth indicator tube 960 system (MGIT) cultures as Mycobacterium tuberculosis (MTB) or NTM together with INNO LiPA Mycobacteria v2 assay (LiPA) and/or PCR sequencing of rDNA for species-specific identification of selected MTB and all NTM isolates in Kuwait. Materials and Methods: DNA was extracted from MGIT cultures (n = 1,033) grown from 664 pulmonary and 369 extrapulmonary specimens from 1,033 suspected tuberculosis patients. mPCR was performed to differentiate MTB from NTM. LiPA was performed and results were interpreted according to kit instructions. rDNA was amplified and sequenced by using panmycobacterial primers. Results: mPCR identified 979 isolates as MTB, 53 as NTM and 1 isolate as mixed culture. LiPA and/or PCR sequencing confirmed 112 of 979 selected isolates as MTB. Mixed culture contained M. tuberculosis and M. fortuitum. LiPA yielded 12 patterns and identified 10 species/species complexes among 47 NTM, M. kansasii + M. scrofulaceum in one culture and 5 isolates only at genus level. PCR sequencing yielded more specific identification for 22 isolates at the species/subspecies level. Conclusions: mPCR rapidly differentiated MTB from NTM. LiPA identified 44 of 52 NTM isolates at the species/species complex level and 2 mixed cultures. PCR sequencing yielded more specific identification at the species/subspecies level. Rapid differentiation as MTB or NTM by mPCR, followed by species-specific NTM identification by LiPA/PCR sequencing is suitable for the proper management of mycobacterial infections in Kuwait.

show abstract

Rapid differentiation of mycobacteria by simplex real-time PCR with melting temperature calling analysis

Abstract: Rapid differentiation of mycobacteria could shorten the diagnostic time of mycobacterial diseases. It is also helpful for achieving optimal therapy and appropriate patient management.

Cited by 2 publications

References 26 publications

Comparison of the Three Molecular Diagnostic Assays for Molecular Identification ofMycobacterium tuberculosisand Nontuberculous Mycobacteria Species in Sputum Samples

Comparison of the Three Molecular Diagnostic Assays for Molecular Identification ofMycobacterium tuberculosisand Nontuberculous Mycobacteria Species in Sputum Samples

Diversity of Nontuberculous Mycobacteria in Kuwait: Rapid Identification and Differentiation of <b><i>Mycobacterium</i></b> Species by Multiplex PCR, INNO-LiPA Mycobacteria v2 Assay and PCR Sequencing of rDNA

Contact Info

Product

Resources

About