聚合酶链反应（PCR）对检测游走性红斑和莱姆神经疏螺旋体病患者临床标本中伯氏疏螺旋体DNA的诊断价值。
Diagnostic value of PCR for detection of Borrelia burgdorferi DNA in clinical specimens from patients with erythema migrans and Lyme neuroborreliosis.
作者信息
Lebech A M, Hansen K, Brandrup F, Clemmensen O, Halkier-Sørensen L
机构信息
Borrelia Laboratory, Department of Clinical Biochemistry, Statens Seruminstitut, Copenhagen.
出版信息
Mol Diagn. 2000 Jun;5(2):139-50. doi: 10.1007/BF03262032.
BACKGROUND
The aim of the study is to evaluate the diagnostic sensitivity of a 16S ribosomal RNA-based PCR on clinical specimens from patients with erythema migrans (EM) and neuroborreliosis and to compare the sensitivities with those obtained by in vitro culture and serological testing. A semiquantitative detection system, representing the input amount of specific DNA and thus the density of spirochetes in clinical specimens, indicated the preferred clinical sample to obtain for PCR testing.
METHODS AND RESULTS
Skin biopsy and urine samples from 31 patients with EM and cerebrospinal fluid (CSF) and urine samples from 30 patients with neuroborreliosis were investigated. Borrelia burgdorferi DNA was detected in 71% of the skin biopsy specimens and 13% of the urine samples from patients with EM. Forty-one percent of the patients with EM were found to have B burgdorferi-specific antibodies in serum, and B burgdorferi was cultured in 29% of the EM specimens. For patients with neuroborreliosis, the diagnostic sensitivities in CSF and urine samples were 17% and 7%, respectively. Specific intrathecal antibody production was found in 90% of the patients, and 87% showed elevated B burgdorferi antibodies in serum. In general, PCR of skin biopsy samples yielded very high amounts of amplicons versus low amounts for CSF and urine samples.
CONCLUSIONS
PCR of skin biopsy specimens is currently the most sensitive and specific test for the diagnosis of patients with EM, superior to culture and serological testing. For B burgdorferi-specific CSF disgnosis in patients with neuroborreliosis, the measurement of specific intrathecal antibody synthesis is superior to PCR. However, in patients with a short duration of disease (<14 days), PCR may be a useful diagnostic supplement. PCR of urine samples cannot be recommended at the present time for routine diagnosis of patients with EM or neuroborreliosis.
背景
本研究的目的是评估基于16S核糖体RNA的聚合酶链反应（PCR）对游走性红斑（EM）和神经型莱姆病患者临床标本的诊断敏感性，并将其敏感性与体外培养和血清学检测所获得的敏感性进行比较。一种半定量检测系统可代表特定DNA的输入量，进而反映临床标本中螺旋体的密度，该系统指明了进行PCR检测时首选的临床样本。
方法与结果
对31例EM患者的皮肤活检和尿液样本以及30例神经型莱姆病患者的脑脊液（CSF）和尿液样本进行了研究。在EM患者的皮肤活检标本中，71%检测到伯氏疏螺旋体DNA，尿液样本中13%检测到该DNA。发现41%的EM患者血清中有伯氏疏螺旋体特异性抗体，29%的EM标本培养出伯氏疏螺旋体。对于神经型莱姆病患者，CSF和尿液样本的诊断敏感性分别为17%和7%。90%的患者发现有特异性鞘内抗体产生，87%的患者血清中伯氏疏螺旋体抗体升高。总体而言，皮肤活检样本的PCR产生的扩增子数量非常多，而CSF和尿液样本的扩增子数量较少。
结论
目前，皮肤活检标本的PCR是诊断EM患者最敏感和特异的检测方法，优于培养和血清学检测。对于神经型莱姆病患者伯氏疏螺旋体特异性CSF诊断，特异性鞘内抗体合成的检测优于PCR。然而，对于病程较短（<14天）的患者，PCR可能是一种有用的诊断补充。目前不推荐将尿液样本的PCR用于EM或神经型莱姆病患者的常规诊断。
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