Development of molecular assays for the analysis of genetic relationships of Mycoplasma iowae.

Autor: Buni D; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary., Kovács ÁB; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, Budapest, Hungary., Földi D; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, Budapest, Hungary., Bányai K; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, Budapest, Hungary; University of Veterinary Medicine Budapest, István utca 2., Budapest 1078, Hungary., Bali K; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, Budapest, Hungary., Domán M; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, Budapest, Hungary., Wehmann E; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, Budapest, Hungary., Bradbury J; Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Neston, Wirral CH64 7TE, United Kingdom., Bottinelli M; Mycoplasma Unit, WOAH Reference Laboratory for Avian Mycoplasmosis, SCT-1, Istituto Zooprofilattico Sperimentale delle Venezie, Via Bovolino 1/C, Buttapietra, Verona 37060, Italy., Catania S; Mycoplasma Unit, WOAH Reference Laboratory for Avian Mycoplasmosis, SCT-1, Istituto Zooprofilattico Sperimentale delle Venezie, Via Bovolino 1/C, Buttapietra, Verona 37060, Italy., Stefani E; Mycoplasma Unit, WOAH Reference Laboratory for Avian Mycoplasmosis, SCT-1, Istituto Zooprofilattico Sperimentale delle Venezie, Via Bovolino 1/C, Buttapietra, Verona 37060, Italy., Lysnyansky I; Department of Avian Diseases, Kimron Veterinary Institute, Beit Dagan, Israel., Kovács L; University of Veterinary Medicine Budapest, István utca 2., Budapest 1078, Hungary; Poultry-Care Kft., Lehel út 21., Újszász 5052, Hungary., Grózner D; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary., Gyuranecz M; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; National Laboratory of Infectious Animal Diseases, Antimicrobial Resistance, Veterinary Public Health and Food Chain Safety, Budapest, Hungary; University of Veterinary Medicine Budapest, István utca 2., Budapest 1078, Hungary; MolliScience Kft., Március 15. utca 1, Biatorbágy 2051, Hungary., Kreizinger Z; Veterinary Medical Research Institute, Hungarian Research Network, Hungária körút 21, Budapest 1143, Hungary; MolliScience Kft., Március 15. utca 1, Biatorbágy 2051, Hungary. Electronic address: kreizinger.zsuzsa@vmri.hun-ren.hu.
Jazyk: angličtina
Zdroj: Veterinary microbiology [Vet Microbiol] 2023 Dec; Vol. 287, pp. 109909. Date of Electronic Publication: 2023 Nov 03.
DOI: 10.1016/j.vetmic.2023.109909
Abstrakt: Mycoplasma iowae is a worldwide spread and economically important avian pathogen that mostly infects turkeys. Currently, multi-locus sequence typing (MLST) serves as the gold standard method for strain identification in M. iowae. However, additional robust genotyping methods are required to effectively monitor M. iowae infections and conduct epidemiological investigations. The first aim of this study was to develop genotyping assays with high resolution, that specifically target M. iowae, namely a multiple-locus variable number of tandem-repeats analysis (MLVA) and a core genome multi-locus sequence typing (cgMLST) schema. The second aim was the determination of relationships among a diverse selection of M. iowae strains and clinical isolates with a previous and the newly developed assays. The MLVA was designed based on the analyses of tandem-repeat (TR) regions in the six serotype reference strains (I, J, K, N, Q and R). The cgMLST schema was developed based on the coding sequences (CDSs) common in 95% of the examined 99 isolates. The samples were submitted for a previously published MLST assay for comparison with the developed methods. Out of 94 TR regions identified, 17 alleles were selected for further evaluation by PCR. Finally, seven alleles were chosen to establish the MLVA assay. Additionally, whole genome sequence analyses identified a total of 676 CDSs shared by 95% of the isolates, all of which were included into the developed cgMLST schema. The MLVA discriminated 19 distinct genotypes (GT), while with the cgMLST assay 79 sequence types (ST) could be determined with Simpson's diversity indices of 0.810 (MLVA) and 0.989 (cgMLST). The applied assays consistently identified the same main clusters among the diverse selection of isolates, thereby demonstrating their suitability for various genetic analyses and their ability to yield congruent results.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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