DETECTION OF BORRELIA-BURGDORFERI IN URINE SPECIMENS FROM DOGS BY A NESTED POLYMERASE-CHAIN-REACTION

A nested PCR (nested flagellin PCR) carrying an internal E-coli DNA co ntrol was established and compared with an in-vitro culture method for the detection of Borrelia burgdorferi in urine specimens of dogs. The predicted specific amplicon of the flagellin gene fla was generated f rom all cultured strains of B-burgdorferi tested (comprising three Eur opean genospecies). In contrast, all 13 strains of seven other flagell ated feri per ml of double-distilled water and approx. 250 bacteria pe r ml of dog urine. Using the bacterial culture method, urine specimens collected from 216 dogs in Germany were all diagnosed negative for sp irochetes by in-vitro culture and dark-field microscopy. In contrast, DNA of B-burgdorferi was detected in 32 specimens (14.8%) by PCR. 31 u rine specimens (14.4) showed inhibitory activity in the PCR assay. How ever, 94 (44%) were inhibitory in the culture assay. The majority of t he PCR-positive dogs exhibited major clinical symptoms which have not been reported in the course of B-burgdorferi infection previously, e.g . cystitis (14/32 dogs) or prostatitis (5/32 dogs). Our results indica te that the analysis of urine specimens by the nested flagellin PCR is a highly valuable procedure for the diagnosis of B-burgdorferi infect ions in dogs.