Construction of a Recombinant Plasmid as Reaction Control in Routine PCR for Detection of Contagious Equine Metritis (CEM-PCR)

Journal of Veterinary Medical Science

Hidekazu NIWA¹  Toru ANZAI¹  Seiji HOBO¹ 

[1] Epizootic Research Center, Equine Research Institute, Japan Racing Association

关键词: reaction control;    PCR;    Taylorella equigenitalis;   

DOI  :  10.1292/jvms.69.1199

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(12)Cited-By(3)Contagious equine metritis (CEM) is a highly contagious bacterial venereal disease of horses caused by Taylorella equigenitalis. CEM-PCR is a semi-nested PCR method for detecting this bacterium. Although this technique is regarded as a sensitive diagnostic method for CEM, there are risks of it generating false positive and false negative results. In this study, we constructed a recombinant plasmid (CEM-POS) as reaction control to assure adequate PCR reaction and prevent false positive results caused by contamination of the reaction control in routine CEM-PCR examinations. CEM-POS was constructed by insertion of rpoB fragments from Rhodococcus equi into CEM-1P, which is a recombinant plasmid that includes a T. equigenitalis-specific sequence region. In CEM-PCR, the size of the PCR product from CEM-POS was clearly different from the true positive PCR product. In addition, CEM-POS retained high stability under convenient storage conditions of 4°C. These results suggest CEM-POS to be a useful tool as a reaction control in routine CEM-PCR examinations.

【 授权许可】

Unknown