Rapid and sensitive detection of waterfowl mycoplasmas using TaqMan assays

Edina Nemesházi; Enikő Wehmann; Dénes Grózner; Dorottya Sára Nagy; Áron Botond Kovács; Dorottya Földi; Zsuzsa Kreizinger; Miklós Gyuranecz

doi:10.1371/journal.pone.0288066

Rapid and sensitive detection of waterfowl mycoplasmas using TaqMan assays

PLoS One. 2023 Jul 14;18(7):e0288066. doi: 10.1371/journal.pone.0288066. eCollection 2023.

Authors

Edina Nemesházi¹, Enikő Wehmann¹, Dénes Grózner¹, Dorottya Sára Nagy¹, Áron Botond Kovács¹, Dorottya Földi¹, Zsuzsa Kreizinger^{1

2}, Miklós Gyuranecz^{1

2

3}

Affiliations

¹ Veterinary Medical Research Institute, Eötvös Loránd Research Network, Budapest, Hungary.
² MolliScience Kft., Biatorbágy, Hungary.
³ University of Veterinary Medicine Budapest, Budapest, Hungary.

Abstract

Waterfowl-specific mycoplasmas cause significant economic losses worldwide. However, only limited resources are available for the specific detection of three such bacteria, Mycoplasma anatis, M. anseris and M. cloacale. We developed species-specific TaqMan assays and tested their reliability across 20 strains of the respective target species as well as 84 non-target avian bacterial strains. Furthermore, we analysed 32 clinical DNA samples and compared the results with those of previously published conventional PCRs. The TaqMan assays showed 100% specificity and very high sensitivity, enabling the detection of target DNA as low as either 10 or 100 copies/μl concentration, depending on the assay. Importantly, we found that while the here developed TaqMan assays are reliable for species-specific detection of M. anatis, the previously published conventional PCR assay may give false positive results. In conclusion, the new assays are reliable, sensitive and suitable for clinical diagnostics of the target species.

Copyright: © 2023 Nemesházi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

MeSH terms

Animals
Birds*
Mycoplasma Infections* / diagnosis
Mycoplasma Infections* / microbiology
Mycoplasma Infections* / veterinary
Polymerase Chain Reaction / methods
Reproducibility of Results
Sensitivity and Specificity

Grants and funding

This work was supported by the KKP19-129751 (MG) and FK21-137809 (ZK) grants of the National Research, Development and Innovation Office (https://nkfih.gov.hu/palyazoknak), Hungary, the SA-27/2021 (MG) grant of the Eötvös Loránd Research Network (https://elkh.org/en), the Project no. RRF-2.3.1-21-2022-00001 (MG) which has been implemented with the support provided by the Recovery and Resilience Facility (RRF), financed under the National Recovery Fund budget estimate, RRF-2.3.1-21 funding scheme and the support provided by the Ministry of Innovation and Technology of Hungary (legal successor: Ministry of Culture and Innovation of Hungary) from the National Research, Development and Innovation Fund, financed under the TKP2021-EGA-01 (MG) funding scheme of the National Research, Development and Innovation Office. Prepared with the professional support of the Doctoral Student Scholarship Program (KÁB, DF) of the Co-operative Doctoral Program of the Ministry of Innovation and Technology, financed from the National Research, Development and Innovation Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.