Bauerfeind R, Kreis U, Weiss R, Wieler L H, Baljer G
Institut für Hygiene und Infektionskrankheiten der Tiere, Justus-Liebig-Universität Giessen, Germany.
Zentralbl Bakteriol. 1998 May;287(4):347-61. doi: 10.1016/s0934-8840(98)80170-9.
A nested PCR (nested flagellin PCR) carrying an internal E. coli DNA control 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 from all cultured strains of B. burgdorferi tested (comprising three European genospecies). In contrast, all 13 strains of seven other flagellated bacterial species were negative. The PCR detection limit yielded 20 cells of B. burgdorferi per ml of double-distilled water and approx. 250 bacteria per ml of dog urine. Using the bacterial culture method, urine specimens collected from 216 dogs in Germany were all diagnosed negative for spirochetes by in-vitro culture and dark-field microscopy. In contrast, DNA of B. burgdorferi was detected in 32 specimens (14.8%) by PCR. 31 urine specimens (14.4%) showed inhibitory activity in the PCR assay. However, 94 (44%) were inhibitory in the culture assay. The majority of the 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 indicate that the analysis of urine specimens by the nested flagellin PCR is a highly valuable procedure for the diagnosis of B. burgdorferi infections in dogs.
建立了一种带有内部大肠杆菌DNA对照的巢式PCR（巢式鞭毛蛋白PCR），并将其与体外培养方法进行比较，用于检测犬尿液标本中的伯氏疏螺旋体。从所有测试的伯氏疏螺旋体培养菌株（包括三种欧洲基因型）中产生了鞭毛蛋白基因fla的预测特异性扩增子。相比之下，其他七种鞭毛细菌的所有13个菌株均为阴性。PCR检测限为每毫升双蒸水中20个伯氏疏螺旋体细胞，每毫升犬尿液中约250个细菌。使用细菌培养方法，对从德国216只犬采集的尿液标本进行体外培养和暗视野显微镜检查，所有标本均诊断为螺旋体阴性。相比之下，通过PCR在32个标本（14.8%）中检测到伯氏疏螺旋体DNA。31个尿液标本（14.4%）在PCR检测中显示出抑制活性。然而，94个（44%）在培养检测中具有抑制作用。大多数PCR阳性犬表现出以前在伯氏疏螺旋体感染过程中未报告的主要临床症状，例如膀胱炎（14/32只犬）或前列腺炎（5/32只犬）。我们的结果表明，通过巢式鞭毛蛋白PCR分析尿液标本是诊断犬伯氏疏螺旋体感染的一种非常有价值的方法。
