Brettschneider S, Bruckbauer H, Klugbauer N, Hofmann H
Klinik für Dermatologie und Allergologie am Biederstein, Technische Universität München, Munich, Germany.
J Clin Microbiol. 1998 Sep;36(9):2658-65. doi: 10.1128/JCM.36.9.2658-2665.1998.
Skin biopsies of 36 patients with erythema migrans and acrodermatitis chronica atrophicans (ACA) before therapy and those of 8 patients after therapy were examined for Borrelia burgdorferi DNA by PCR. Skin biopsies of 27 patients with dermatological diseases other than Lyme borreliosis and those of 10 healthy persons were examined as controls. Two different primer sets targeting 23S rRNA (PCR I) and 66-kDa protein (PCR II) genes were used. PCR was performed with freshly frozen tissue (FFT) and paraffin-embedded tissue (PET). For FFT specimens of erythema migrans, 73% were positive by PCR I, 79% were positive by PCR II, and 88% were positive by combining PCR I and II. For PET specimens, PCR was less sensitive (PCR I, 44%; PCR II, 52%). For FFT specimens of ACA, PCR I was positive for two of five patients and PCR II was positive for four of five patients. B. burgdorferi was cultured from 79% of the erythema migrans specimens but not from any of the ACA lesions. Elevated B. burgdorferi antibodies were detected in sera of 74% of erythema migrans patients and 100% of ACA patients. All urine samples were negative by PCR II, whereas PCR I was positive for 27%. However, hybridization of these amplicons was negative. Sequencing of three amplicons identified nonborrelial DNA. In conclusion, urine PCR is not suitable for the diagnosis of skin borreliosis. A combination of two different primer sets achieves high sensitivity with skin biopsies. In early erythema migrans infection, culture and PCR are more sensitive than serology.
对36例游走性红斑和慢性萎缩性肢端皮炎(ACA)患者治疗前的皮肤活检样本以及8例治疗后的皮肤活检样本进行PCR检测,以查找伯氏疏螺旋体DNA。对27例非莱姆病螺旋体病的皮肤病患者以及10名健康人的皮肤活检样本进行检测作为对照。使用了两种针对23S rRNA(PCR I)和66-kDa蛋白(PCR II)基因的不同引物组。PCR检测采用新鲜冷冻组织(FFT)和石蜡包埋组织(PET)。对于游走性红斑的FFT样本,PCR I的阳性率为73%,PCR II的阳性率为79%,将PCR I和II结合使用时阳性率为88%。对于PET样本,PCR的敏感性较低(PCR I为44%;PCR II为%)。对于ACA的FFT样本,5例患者中有2例PCR I呈阳性,5例患者中有4例PCR II呈阳性。79%的游走性红斑样本培养出伯氏疏螺旋体,但ACA病变样本均未培养出。74%的游走性红斑患者血清和100%的ACA患者血清检测到伯氏疏螺旋体抗体升高。所有尿液样本PCR II检测均为阴性,而PCR I的阳性率为27%。然而,这些扩增子的杂交检测为阴性。对三个扩增子进行测序鉴定出非疏螺旋体DNA。总之,尿液PCR不适用于皮肤疏螺旋体病的诊断。两种不同引物组结合使用可使皮肤活检具有高敏感性。在游走性红斑早期感染中,培养和PCR比血清学检测更敏感。