20.500.12556/DiRROS-14710 Next-generation sequencing to characterize pyrazinamide resistance in Mycobacterium tuberculosis isolates from two Balkan countries 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. tuberculosis Mycobacterium tuberculosis high-throughput nucleotide sequencing pyrazinamide microbial sensitivity tests next-generation sequencing drug susceptibility testing Slovenia Republic of North Macedonia tuberkuloza Mycobacterium tuberculosis visokopretočno sekvenciranje pirazinamid test občutljivosti mikroorganizmov sekvenciranje naslednje generacije ugotavljanje občutljivosti za zdravila Slovenija Republika Severna Makedonija true false true Elsevier on behalf of International Society for Antimicrobial Chemotherapy Angleški jezik Ni določen © 2021 The Authors Neznano 2022-01-10 13:32:41 2022-01-10 13:32:43 2022-08-20 03:37:21 0000-00-00 00:00:00 2021 0 Nizozemska 0 Nasl. z nasl. zaslona; Soavtorji: Simon Koren, Nataša Toplak, Sara Truden, Manca Žolnir-Dovč; Opis vira z dne 15. 12. 2021; str. 1-25 Vol. 2021 0000-00-00 PostprintKoncna VTisku NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616-002 2213-7173 10.1016/j.jgar.2021.09.019 89580803 RAZ_Sodja_Eva_i2021.pdf RAZ_Sodja_Eva_i2021.pdf 1 95947C58B237E922AB6302C82F389C72 3fb0f9f72c514898f9ab1ae92b3661c4d1efa0eb6ea62376874f416047bf510e cf1b570a-17b6-11ed-b6b8-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=18899 https://www.sciencedirect.com/science/article/pii/S221371652100254X/pdfft?md5=fb191770c5ca327e3f1758114e724d42&pid=1-s2.0-S221371652100254X-main.pdf 1 https://dirros.openscience.si/Dokument.php?lang=slv&id=18898 Univerzitetna klinika za pljučne bolezni in alergijo Golnik 0 0 0