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Updated: Jun 29, 2026
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
A real-time polymerase chain reaction assay for the detection of Mycoplasma agalactiae
J Fitzmaurice1, M Sewell, C M King
1Investigation and Diagnostic Centre, Biosecurity New Zealand, Ministry of Agriculture and Forestry, Upper Hutt, New Zealand.
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.
