Appuhamy S, Low J C, Parton R, Coote J G
Division of Infection and Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, UK.
J Appl Microbiol. 1998 Dec;85(6):941-8. doi: 10.1111/j.1365-2672.1998.tb05257.x.
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.
精液放线杆菌是绵羊附睾炎和公羊不育症的常见病因。对于公羊精液样本的质量控制而言，迅速检测和鉴定这种微生物在商业上具有重要意义。精液放线杆菌是一种苛求且生长缓慢的细菌，初次分离和初步鉴定可能既困难又耗时。在本研究中，已对精液放线杆菌基因组中的两个核糖体操纵子（称为rrnA和rrnB）进行了表征，它们在16S和23S rRNA基因之间的间隔区分别含有一个和两个tRNA。根据rrnB间隔区序列开发了精液放线杆菌的种特异性引物，用于通过PCR鉴定和检测精液放线杆菌。该PCR检测方法对精液放线杆菌具有特异性，对表型相似的微生物（如绵羊嗜组织菌）未产生扩增产物。发现用于长期保存精液的保存液会抑制PCR。因此，出于诊断目的，该检测最好在初次分离后进行，或者如果有明显污染迹象，也可以在精液保存前对新鲜精液进行检测。
