Comparison of polymerase chain reaction and cell culture for the detection of Chlamydophila species in the semen of bulls, buffalo-bulls, and rams

Two hundred and thirty six semen samples were collected from 120 bulls, 60 buffalo-bulls, and 56 rams located on farms of known history of infection with Chlamydophila species. All semen samples were examined by polymerase chain reaction (PCR) and cell culture techniques for detection of Chlamydophila species. The primers were selected to allow the amplification of all target species in a single reaction by identifying conserved sequences in the omp2 gene. PCR assay detected more positive samples (36) from the semen samples collected from different animal species than were detected by the culture method (21). The results indicated that all culture-positive semen samples (21) from different species were PCR positive. The detection limit of the PCR assay was determined with DNA extracted from fourfold serial dilution of C. abortus (B577) and C. pecorum (11/88) cultures and found to be 0.25 inclusion-forming units (IFU) per PCR, while the culture method could not detect less than 4 IFU. This is the first report using PCR for the detection of Chlamydophila species in buffalo-bulls’ semen and the assay provides a simple, sensitive, rapid, and reliable means for the detection and identification of the organism.