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Title:Next-generation sequencing to characterize pyrazinamide resistance in Mycobacterium tuberculosis isolates from two Balkan countries
Authors:ID Sodja, Eva, Klinika Golnik (Author)
ID Koren, Simon (Author)
ID Toplak, Nataša (Author)
ID Truden, Sara, Klinika Golnik (Author)
ID Žolnir-Dovč, Marija, Klinika Golnik (Author)
Files:.pdf PDF - Presentation file, download (1,53 MB)
MD5: 95947C58B237E922AB6302C82F389C72
URL URL - Source URL, visit
Typology:1.01 - Original Scientific Article
Organization:Logo UKPBAG - University Clinic of Respiratory and Allergic Diseases Golnik
Abstract:Objectives. Next-generation sequencing (NGS) provide a comprehensive analysis of the genetic alterations that are most commonly linked with pyrazinamide (PZA) resistance. There are no studies reporting molecular background of PZA resistance in TB isolates from Balkan Peninsula. We aimed to examine the feasibility of full-length analysis of a gene linked with PZA resistance, pncA, using Ion Torrent technology in comparison to phenotypic BACTEC MGIT 960 DST in clinical TB isolates from two countries of the Balkan Peninsula. Methods. Between 1996 and 2017, we retrospectively selected 61 TB isolates. To identify gene variants related to drug resistance in genomic DNA extracted from TB isolates, AmpliSeq libraries were generated automatically using the AmpliSeq™ Kit for Chef DL8 and the Ion AmpliSeq TB Research Panel. Result.s Of all 61 TB isolates included, 56 TB were phenotypically resistant to any antibiotic. Among them, 38/56 (67.9%) TB isolates were phenotypically resistant to pyrazinamide and pncA mutations were detected in 33/38 cases (86.8%). A mutation in the pncA promoter region was the most prevalent genetic alteration, detected in eight TB isolates. Comparison of NGS to conventional BACTEC MGIT 960 DST revealed very strong agreement (90.2%) between the two methods in identifying PZA resistance, with high sensitivity (89.5%) and specificity (95.7%) for NGS. Conclusions. Detection of PZA resistance using NGS seems to be a valuable tool for surveillance of TB drug resistance also in the Balkan Peninsula, with great potential to provide useful information at least one weak earlier than is possible with phenotypic DST.
Keywords:tuberculosis, Mycobacterium tuberculosis, high-throughput nucleotide sequencing, pyrazinamide, microbial sensitivity tests, next-generation sequencing, drug susceptibility testing, Slovenia, Republic of North Macedonia
Year of publishing:2021
Publication status in journal:In print
Article version:Postprint, final article version, accepted into publication
Publisher:Elsevier on behalf of International Society for Antimicrobial Chemotherapy
Number of pages:str. 1-25
ISSN on article:2213-7173
DOI:10.1016/j.jgar.2021.09.019 New window
COBISS.SI-ID:89580803 New window
Copyright:© 2021 The Authors
Note:Nasl. z nasl. zaslona; Soavtorji: Simon Koren, Nataša Toplak, Sara Truden, Manca Žolnir-Dovč; Opis vira z dne 15. 12. 2021;
Publication date in DiRROS:10.01.2022
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Record is a part of a journal

Title:Journal of global antimicrobial resistance
COBISS.SI-ID:56412419 New window


License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:21.11.2021

Secondary language

Keywords:tuberkuloza, Mycobacterium tuberculosis, visokopretočno sekvenciranje, pirazinamid, test občutljivosti mikroorganizmov, sekvenciranje naslednje generacije, ugotavljanje občutljivosti za zdravila, Slovenija, Republika Severna Makedonija