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通过聚合酶链反应检测早期和晚期莱姆病神经伯氏疏螺旋体病患者尿液样本和脑脊液样本中的伯氏疏螺旋体DNA。

Detection of Borrelia burgdorferi DNA in urine samples and cerebrospinal fluid samples from patients with early and late Lyme neuroborreliosis by polymerase chain reaction.

作者信息

Lebech A M, Hansen K

机构信息

Department of Infection-Immunology, Statens Seruminstitut, Copenhagen, Denmark.

出版信息

J Clin Microbiol. 1992 Jul;30(7):1646-53. doi: 10.1128/jcm.30.7.1646-1653.1992.

Abstract

A polymerase chain reaction (PCR) was developed for use in the identification of a 248-bp fragment of the Borrelia burgdorferi flagellin gene in urine and cerebrospinal fluid (CSF) from patients with Lyme neuroborreliosis. The specificities of the PCR products were confirmed by DNA-DNA hybridization with an internal probe. The assay had a detection limit of 10 in vitro-cultivated B. burgdorferi. The PCR assay seemed to be species wide as well as species specific, since DNA from all 21 B. burgdorferi isolates from humans tested but not from Borrelia hermsii or Treponema pallidum could be amplified. We tested 10 consecutively diagnosed patients with untreated neuroborreliosis. There was lymphocytic pleocytosis and intrathecal B. burgdorferi-specific antibody synthesis in the CSF of all patients. Urine and CSF samples were investigated by PCR before, during, and up to 8.5 months after therapy. B. burgdorferi DNA was detected in urine samples from nine patients; five patients, including two patients with chronic neuroborreliosis, were PCR positive prior to treatment, whereas urine samples from the remaining four patients obtained 3 to 6 days after the onset of therapy became PCR positive. All urine samples obtained greater than 4 weeks after therapy were negative by PCR. PCR of CSF was less sensitive, and samples from only four patients, including one with chronic neuroborreliosis, were positive. We conclude that urine is a more suitable sample source than CSF for use in B. burgdorferi DNA detection by PCR. Normalization of inflammatory CSF changes and the negative PCR results during follow-up even in patients with chronic neuroborreliosis do not point to a persistent infection. The future role of PCR as a diagnostic tool for Lyme neuroborreliosis is still uncertain.

摘要

开发了一种聚合酶链反应(PCR),用于鉴定莱姆神经疏螺旋体病患者尿液和脑脊液(CSF)中伯氏疏螺旋体鞭毛蛋白基因的248 bp片段。通过与内部探针进行DNA-DNA杂交来确认PCR产物的特异性。该检测方法对体外培养的伯氏疏螺旋体的检测限为10个菌。该PCR检测方法似乎具有种属广泛性和种属特异性,因为来自所有21株检测的人类伯氏疏螺旋体分离株的DNA均可扩增,但赫氏疏螺旋体或梅毒螺旋体的DNA则不能扩增。我们对10例连续诊断的未经治疗的神经疏螺旋体病患者进行了检测。所有患者的脑脊液中均有淋巴细胞增多和鞘内伯氏疏螺旋体特异性抗体合成。在治疗前、治疗期间以及治疗后长达8.5个月对尿液和脑脊液样本进行PCR检测。在9例患者的尿液样本中检测到伯氏疏螺旋体DNA;5例患者,包括2例慢性神经疏螺旋体病患者,在治疗前PCR呈阳性,而其余4例患者在治疗开始后3至6天采集的尿液样本PCR呈阳性。治疗后超过4周采集的所有尿液样本PCR均为阴性。脑脊液的PCR检测敏感性较低,只有4例患者的样本呈阳性,其中包括1例慢性神经疏螺旋体病患者。我们得出结论,对于通过PCR检测伯氏疏螺旋体DNA,尿液是比脑脊液更合适的样本来源。即使是慢性神经疏螺旋体病患者,脑脊液炎症变化的正常化以及随访期间PCR阴性结果并不表明存在持续性感染。PCR作为莱姆神经疏螺旋体病诊断工具的未来作用仍不确定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad71/265358/2ce50dff4e70/jcm00031-0047-a.jpg

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