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Abstract
AIM: To develop a real-time PCR for the detection of Mycoplasma agalactiae, using PCR primers targeting the ma-mp81 gene.
METHODS: A group of 15 M. agalactiae isolates, 21 other Mycoplasma spp. isolates and 21 other bacterial isolates was used in evaluation of the assay.
RESULTS: All M. agalactiae isolates were detected by the assay and none of the non-target isolates was amplified. The analytical detection limit of the assay was 10 fg of purified genomic DNA and 104 cfu/ml milk inoculated with M. agalactiae. When applied to goat-milk samples collected from three herds free of M. agalactiae infection, the assay had a specificity of 100%.
CONCLUSIONS: The assay would be useful in a diagnostic laboratory, providing specific, sensitive and rapid detection of M. agalactiae.
KEY WORDS:
Acknowledgements
The authors gratefully acknowledge the kind donation of DNA from Dr Marc Marenda (University of Melbourne, Australia), Dr Göran Bölske (National Veterinary Institute, Sweden), Mr Trevor Taylor (AAHL, Australia), Dr François Thiaucourt (CIRAD, France), and Professor Joachim Frey (Institute of Veterinary Bacteriology, Switzerland). Fiona Anniss is thanked for collection of milk samples from goats.