Pyrosequencing for rapid detection of tuberculosis resistance to Rifampicin and Isoniazid in Syrian and Lebanese clinical isolates. Issue 3 (September 2015)

• Record Type: Journal Article
• Authors: Hamze, M.; Ismail, M.B.; Rahmo, A.K.; Dabboussi, F.
• Is Part Of: International journal of mycobacteriology. Volume 4:Issue 3(2015:Sep.)
• Page Start: 228
• Page End: 232
• Publication Date: 2015-09
• Subjects: Mycobacteria -- Periodicals; Mycobacterial diseases -- Periodicals; Mycobacteriaceae; Mycobacteria; Electronic journals; Periodicals; 579.374
• DOI: 10.1016/j.ijmyco.2015.05.007
• Languages: English
• ISSNs: 2212-5531

Abstract

<abstract xml:lang="en" abstract-type="author" id="ab005"> <title id="st005">Abstract</title> <sec> <title id="st010">Background</title> <p id="sp0005">Rapid and accurate techniques are always welcomed for the detection of resistant strains of <italic>Mycobacterium tuberculosis</italic> MTB.</p> </sec> <sec> <title id="st015">Objectives</title> <p id="sp0010">The objective of this study is to evaluate the pyrosequencing technology for the detection of MTB resistance to Rifampicin (RIF) and Isoniazid (INH) in Syrian and Lebanese clinical strains; 66 strains resistant to INH, among them 56 resistant also to RIF, were tested.</p> </sec> <sec> <title id="st020">Methods</title> <p id="sp0015">Four pyrosequencing assays were optimized and applied to the following loci: <italic>rpoB</italic>rpoB RIF resistance-determining region, <italic>katG</italic>, the promoter regions of <italic>inhA</italic> and <italic>ahpC</italic>-<italic>oxyR</italic> intergenic region.</p> </sec> <sec> <title id="st025">Results</title> <p id="sp0020">The prevalence of mutations on codon 315 of the <italic>katG</italic> gene, <italic>inhA</italic> and <italic>ahpc</italic>-<italic>oxyR</italic> were 42.4%, 21.2% and 9.0%, respectively, which make an overall sensitivity of 72.6% for INH resistance. All RIF-resistant strains contained at least one non-synonymous codon change in the sequenced <italic>rpoB</italic> region (507–533) relative to the ATCC reference strain. The RIF drug resistance region (RRDR) sequencing identified 96 modified codons representing 34 different mutations.</p> </sec> <sec> <title id="st030">Conclusions</title> <p id="sp0025">The high sensitivity and the short turnaround time combined with multilocus sequencing of several isolates in parallel make pyrosequencing an attractive method for drug resistance screening for MTB<italic>.</italic></p> </sec> </abstract>