Specific PCR primers from the 16S-23S rRNA spacer region for the rapid detection and identification of Actinobacillus seminis.
Actinobacillus seminis is a common cause of ovine epididymitis and ram infertility. The ability to detect and identify this organism promptly is important commercially for the quality control of ram semen samples. Actinobacillus seminis is a fastidious and slow-growing bacterium and primary isolation and presumptive identification can be difficult and time-consuming. In this study, two ribosomal operons, termed rrnA and rrnB, have been characterized in the A. seminis genome, and these contain one and two tRNAs, respectively, in the spacer region between the 16S and 23S rRNA genes. Species-specific primers for A. seminis were developed from the sequence of the spacer region of rrnB for the identification and detection of A. seminis by PCR. The PCR assay was specific for A. seminis and gave no amplification products with phenotypically similar organisms such as Histophilus ovis. Storage solution used to preserve semen for long-term storage was found to inhibit the PCR. Therefore, for diagnostic purposes, the assay would best be performed after primary isolation or perhaps on fresh semen prior to storage if obvious contamination is indicated.
Appuhamy
S
S
Division of Infection and Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, UK.
Low
J C
JC
Parton
R
R
Coote
J G
JG
D000188:Actinobacillus / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000032:analysis*; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D012661:Semen / Q000382:microbiology; D012680:Sensitivity and Specificity; D012756:Sheep; D013997:Time Factors; D012407:rRNA Operon
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S