Detection of Mycoplasma anatis, M. anseris, M. cloacale and Mycoplasma sp. 1220 in waterfowl using species-specific PCR assays.

Autor: Grózner D; Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary., Sulyok KM; Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary., Kreizinger Z; Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary., Rónai Z; Veterinary Diagnostic Directorate, National Food Chain Safety Office, Budapest, Hungary., Jánosi S; Veterinary Diagnostic Directorate, National Food Chain Safety Office, Budapest, Hungary., Turcsányi I; Veterinary Diagnostic Directorate, National Food Chain Safety Office, Budapest, Hungary., Károlyi HF; Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary.; Department of Microbiology and Infectious Diseases, University of Veterinary Medicine, Budapest, Hungary., Kovács ÁB; Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary., Kiss MJ; TOLL 96 Kft., Hajdúsámson, Hungary., Volokhov D; Center for Biologics Evaluation and Research, U. S. Food and Drug Administration, Silver Spring, Maryland, United States of America., Gyuranecz M; Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary.; Department of Microbiology and Infectious Diseases, University of Veterinary Medicine, Budapest, Hungary.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2019 Jul 11; Vol. 14 (7), pp. e0219071. Date of Electronic Publication: 2019 Jul 11 (Print Publication: 2019).
DOI: 10.1371/journal.pone.0219071
Abstrakt: Mycoplasma anatis, M. anseris, M. cloacale and M. sp. 1220 colonise geese and ducks, and could be associated with infections of avian respiratory and nervous systems, cause mild to severe inflammation of cloaca and genital tracts, and embryo lethality. Co-occurrence of these Mycoplasma species in waterfowl is frequently detected and the identification of these mycoplasmas to the species level at a regular microbiology laboratory is difficult due to their similar morphological, cultural and biochemical properties. Moreover, species differentiation is only possible based on the sequence analysis of the product of a genus-specific PCR assay. Therefore, the aim of the current study was to develop an effective and robust method for the identification of these species in avian clinical specimens. Polymerase chain reaction (PCR) assays using species-specific primers, which target housekeeping genes in order to identify these species, were designed in the present study. The developed PCR assays can precisely identify these four mycoplasmas to the species level directly from DNA samples extracted from clinical specimens, and no cross-amplification was observed among these species and with other well-known avian mycoplasmas. The average sensitivity of the assays was 101-102 genomic equivalents per reaction. These conventional PCR assays can be run simultaneously at the same PCR cycling program, and the species can be differentiated directly (without sequence analysis) by gel electrophoresis due to the specific sizes of the amplicons. In conclusion, the presented species-specific assays were found to be suitable for routine use at regular veterinary diagnostic laboratories and promote the rapid, simple and cost-effective differentiation of these waterfowl Mycoplasma species.
Competing Interests: The TOLL 96 Kft. provided support in the form of salaries for MJK. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
Databáze: MEDLINE
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