Development of a multiplex real-time PCR for contagious agalactia diagnosis in small ruminants

Journal of Microbiological Methods
Claire A M BeckerFlorence Tardy

Abstract

Contagious agalactia is an important disease worldwide that affects small ruminants. Clinical manifestations vary from mastitis, pneumonia, arthritis and keratoconjunctivitis to septicemia. Four mycoplasmal etiological agents have been identified: Mycoplasma (M.) agalactiae, M. mycoides subsp. capri, M. capricolum subsp. capricolum and M. putrefaciens. The current procedure for direct diagnosis, recommended by the World Organization for Animal Health, involves the isolation of one or several causative agents from clinical specimens and further time-consuming identification steps. The present study reports the development of a new multiplex real-time PCR (including an internal positive control) that detects all four pathogens simultaneously and distinguishes M. agalactiae from the others. First, intra- and inter-species polymorphisms of the two target house-keeping genes, namely polC and fusA, were analyzed to design primers and probes adapted to the diversity of currently circulating strains. The specificity and the sensitivity of the assay were then challenged and the limit of detection was found to be as low as 6 to 12 copies of the target genes. The assay requires further assessment on clinical specimens but its performances...Continue Reading

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Citations

Sep 10, 2015·Irish Veterinary Journal·J Tatay-DualdeJ Amores
Aug 7, 2014·Veterinary Medicine International·Amit KumarKuldeep Dhama
Jul 17, 2014·Veterinary Medicine International·Sandip ChakrabortyMinakshi Prasad
Jan 27, 2015·Tropical Animal Health and Production·Valsala RekhaInbaraj Sophia
Mar 12, 2019·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Gayle C JohnsonMichael J Calcutt
Feb 5, 2015·The Journal of Veterinary Medical Science·Falong YangHua Yue

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