Krüger W H, Pulz M
Institute of Medical Microbiology and Immunology, University Hospital Eppendorf, Hamburg, Germany.
J Med Microbiol. 1991 Aug;35(2):98-102. doi: 10.1099/00222615-35-2-98.
The polymerase chain reaction (PCR) was used to amplify specific DNA sequences from different clinical isolates of Borrelia burgdorferi and from cerebrospinal fluid (CSF) of two patients with Lyme disease of the central nervous system. The amplification products were separated by polyacrylamide gel electrophoresis and visualised by ethidium bromide staining. The definitive identification of amplified DNA as a part of the B. burgdorferi flagellin gene was achieved by hybridisation to a 40-base oligonucleotide probe complementary to a part of the spirochaetal gene but not to the primers. Attempts to cultivate borreliae from either patient were unsuccessful and one patient had no serological marker in serum or CSF to indicate borreliosis. Clinical symptoms of both patients regressed with antibiotic therapy. The PCR system is a powerful and rapid technique to amplify flagellin gene sequences from CSF of patients with neuroborreliosis. Only one-tenth of the time needed for cultivation was required from CSF sampling to diagnosis. Gene amplification might, for the first time, allow effective monitoring of therapy for patients with Lyme disease of the central nervous system.
采用聚合酶链反应(PCR)从不同的伯氏疏螺旋体临床分离株以及两名患有中枢神经系统莱姆病患者的脑脊液(CSF)中扩增特定的DNA序列。扩增产物通过聚丙烯酰胺凝胶电泳进行分离,并用溴化乙锭染色进行可视化。通过与一个40碱基的寡核苷酸探针杂交,该探针与螺旋体基因的一部分互补但与引物不互补,从而将扩增的DNA明确鉴定为伯氏疏螺旋体鞭毛蛋白基因的一部分。从两名患者中培养疏螺旋体均未成功,且其中一名患者的血清或脑脊液中没有血清学标志物表明存在疏螺旋体病。两名患者的临床症状经抗生素治疗后均有所缓解。PCR系统是一种强大而快速的技术,可从神经型疏螺旋体病患者的脑脊液中扩增鞭毛蛋白基因序列。从脑脊液采样到诊断仅需培养所需时间的十分之一。基因扩增首次可能允许对中枢神经系统莱姆病患者的治疗进行有效监测。