Multiplex PCR for the detection of Brucella ovis, Actinobacillus seminis and Histophilus somni in ram semen.
OBJECTIVE
To develop a multiplex polymerase chain reaction (PCR) assay for the rapid detection of Brucella ovis, Actinobacillus seminis, Histophilus somni in fresh ram semen samples.


METHODS
The multiplex assay was based on the single PCR assays published for the detection of A seminis and B ovis, and the forward primer published for the detection of H somni; an alternative reverse primer for H somni was designed in this study.


METHODS
Culture and PCR of 295 fresh semen samples were carried out.


RESULTS
The multiplex PCR was far more successful in the detection of H somni (45/295) than culture (23/295). A seminis was also detected in more semen samples by multiplex PCR (29/295) than culture (13/295) and B ovis was detected in three samples using both PCR and culture. No amplifications were detected with DNA from a range of bacterial isolates including species associated with epididymitis in rams.


CONCLUSIONS
This PCR could be used as a complementary test, or alternative to culture of ram semen and other biological samples for the detection B ovis, H somni and A seminis.
Saunders
V F
VF
Elizabeth Macarthur Agricultural Institute, New South Wales Department of Primary Industries, Camden, NSW 2570.
Reddacliff
L A
LA
Berg
T
T
Hornitzky
M
M
D044133:Actinobacillus seminis / Q000302:isolation & purification*; D000818:Animals; D042061:Brucella ovis / Q000302:isolation & purification*; D015169:Colony Count, Microbial / Q000662:veterinary; D004269:DNA, Bacterial / Q000032:analysis; D008297:Male; D015661:Pasteurellaceae / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015203:Reproducibility of Results; D012661:Semen / Q000382:microbiology*; D012680:Sensitivity and Specificity; D012756:Sheep
D004269:DNA, Bacterial