A real-time polymerase chain reaction assay for the detection of Mycoplasma agalactiae.
OBJECTIVE
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.
Investigation and Diagnostic Centre, Biosecurity New Zealand, Ministry of Agriculture and Forestry, Upper Hutt, New Zealand.
J
J
Fitzmaurice
M
M
Sewell
C M
CM
King
S
S
McDougall
W L
WL
McDonald
J S
JS
O'Keefe
D000818:Animals; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015511:Goat Diseases / Q000175:diagnosis* / Q000382:microbiology; D006041:Goats; D008892:Milk / Q000382:microbiology; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D045644:Mycoplasma agalactiae / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000382:microbiology; D013045:Species Specificity
D004269:DNA, Bacterial