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|Title:||Development of multiplex PCR assays based on the 16S-23S rRNA internal transcribed spacer for the detection of clinically relevant nontuberculous mycobacteria||Authors:||Ngan, G.J.Y.
Polymerase chain reaction
|Issue Date:||May-2011||Citation:||Ngan, G.J.Y., Ng, L.M., Jureen, R., Lin, R.T.P., Teo, J.W.P. (2011-05). Development of multiplex PCR assays based on the 16S-23S rRNA internal transcribed spacer for the detection of clinically relevant nontuberculous mycobacteria. Letters in Applied Microbiology 52 (5) : 546-554. ScholarBank@NUS Repository. https://doi.org/10.1111/j.1472-765X.2011.03045.x||Abstract:||Aims: To accelerate the identification and differentiation of clinically relevant nontuberculous mycobacteria (NTM) with two sets of multiplex PCR (mPCR) targeting the 16S-23S rRNA internal transcribed spacer (ITS) region for timely patient management. Methods and Results: Two mPCR assays were developed: Slow-Growers (SG) mPCR was used for the detection of slow-growing mycobacteria, which included Mycobacterium avium complex, Mycobacterium kansasii, Mycobacterium gordonae and Mycobacterium xenopi whilst the other mPCR assay labelled as Fast-Growers (FG) mPCR was used for the detection of Mycobacterium fortuitum complex, Mycobacterium abscessus and Mycobacterium chelonae. In these assays, a common forward primer based on a conserved section of the 16S rRNA region was used in conjunction with species-specific reverse primers. The mPCRs were tested against 247 clinical mycobacterial isolates and demonstrated 100% specificity and sensitivity. Identification of the mycobacterial species was also validated by DNA sequencing of the 16S-23S ITS region and when further confirmation was needed, hsp65 sequencing was performed. Conclusions: The mPCR assays could be a potentially useful diagnostic tool for the rapid and accurate identification of clinically relevant NTM. Significance and Impact of the Study: In this study, we looked at the frequency of hospital isolated NTM over the last 5years (2005-2010), and an mPCR targeting the ITS region was developed for NTM species that appeared to be more prevalent in the context of Singapore. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.||Source Title:||Letters in Applied Microbiology||URI:||http://scholarbank.nus.edu.sg/handle/10635/125626||ISSN:||02668254||DOI:||10.1111/j.1472-765X.2011.03045.x|
|Appears in Collections:||Staff Publications|
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