REAL

Development of Molecular Methods for the Rapid Differentiation of Mycoplasma gallisepticum Vaccine Strains from Field Isolates

Sulyok, Kinga Mária and Kreizinger, Zsuzsa and Bekő, Katinka and Forró, Barbara and Marton, Szilvia and Bányai, Krisztián and Catania, Salvatore and Ellis, Christine and Bradbury, Janet and Olaogun, Olusola M and Kovács, Áron Botond and Cserép, Tibor and Gyuranecz, Miklós (2019) Development of Molecular Methods for the Rapid Differentiation of Mycoplasma gallisepticum Vaccine Strains from Field Isolates. Journal of Clinical Microbiology, 57 (6). ISSN 0095-1137 (print), 1098-660X (online)

[img] Text
Development of Molecular Methods for the Rapid differentiation of Mycoplasma gallisepticum vaccine strains from field isolates.pdf - Published Version
Restricted to Repository staff only

Download (1MB) | Request a copy

Abstract

Mycoplasma gallisepticum is among the economically most significant mycoplasmas causing production losses in poultry. Seven melt-curve and agarose gel based mismatch amplification mutation assays (MAMA) and one polymerase chain reaction (PCR) are provided in the present study to distinguish the M. gallisepticum vaccine strains and field isolates based on mutations in crmA, gapA, lpd, plpA, potC, glpK, and hlp2 genes. A total of 239 samples (M. gallisepticum vaccine and type strains, pure cultures and clinical samples) originating from 16 countries and from at least eight avian species were submitted to the presented assays for validation or in blind tests. Comparison of the data of 126 samples (including sequences available at GenBank) examined by the developed assays and a recently developed multi-locus sequence typing assay showed congruent typing results. The sensitivity of melt-MAMA assays varied between 101-104 M. gallisepticum template copy number/reaction, while that of the agarose-MAMAs ranged between 103 and 105 template copy number/reaction and no cross-reactions occurred with other Mycoplasma species colonizing birds. The presented assays are also suitable to discriminate multiple strains in a single sample. The developed assays enable the differentiation of live vaccine strains by targeting two or three markers/vaccine strain; however, considering the high variability of the species, the combined use of all assays is recommended. The suggested combination provides a reliable tool for routine diagnostics, due to the sensitivity and specificity of the assays, and that they can be performed directly on clinical samples and in laboratories with basic PCR equipment.

Item Type: Article
Subjects: S Agriculture / mezőgazdaság > SV Veterinary science / állatorvostudomány
Depositing User: Dr. Enikő Wehmann
Date Deposited: 16 Sep 2019 10:45
Last Modified: 16 Sep 2019 10:45
URI: http://real.mtak.hu/id/eprint/99513

Actions (login required)

Edit Item Edit Item