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Multiplex PCR for the detection of Brucella ovis , Actinobacillus seminis and Histophilus somni in ram semen
Author(s) -
Saunders VF,
Reddacliff LA,
Berg T,
Hornitzky M
Publication year - 2007
Publication title -
australian veterinary journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.382
H-Index - 59
eISSN - 1751-0813
pISSN - 0005-0423
DOI - 10.1111/j.1751-0813.2006.00098.x
Subject(s) - semen , multiplex polymerase chain reaction , microbiology and biotechnology , biology , brucella , polymerase chain reaction , primer (cosmetics) , multiplex , brucellosis , virology , chemistry , genetics , gene , organic chemistry
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. Design 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. Procedure 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. Conclusion 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 .