Liebling M R, Nishio M J, Rodriguez A, Sigal L H, Jin T, Louie J S
Division of Rheumatology, Harbor-University of California, Los Angeles Medical Center, Torrance 90509.
Arthritis Rheum. 1993 May;36(5):665-75. doi: 10.1002/art.1780360514.
To analyze clinical fluids for the presence of Borrelia burgdorferi DNA using the polymerase chain reaction (PCR).
We utilized a modified, nested PCR to detect the presence of Borrelia DNA in 99 samples of serum, urine, cerebrospinal fluid (CSF), or synovial fluid obtained from 44 patients with various stages of Lyme disease and 47 control subjects. Primer specificity was corroborated by examining 2 DNA data banks, testing against DNA from other organisms, and confirming results with a second set of nested primers.
Nested PCR was capable of detecting DNA from fewer than 10 organisms in 1 ml of fluid. The specificity of this technique was 96.4%, with a sensitivity of 76.7%. Although the specificity was uniformly high, the sensitivity was dependent upon the body fluid being tested: CSF 100%, urine 100%, synovial fluid 80%, and serum 59%. The rate of false-positive results was 3.6%.
These data demonstrate the potential utility of PCR in confirming the clinical diagnosis of Lyme disease as well as providing insight into the pathogenesis of various stages of this disorder.
使用聚合酶链反应(PCR)分析临床体液中是否存在伯氏疏螺旋体DNA。
我们采用改良的巢式PCR检测从44例处于莱姆病不同阶段的患者和47名对照受试者获取的99份血清、尿液、脑脊液(CSF)或滑液样本中伯氏疏螺旋体DNA的存在情况。通过检查2个DNA数据库、与其他生物体的DNA进行测试以及用第二组巢式引物确认结果来证实引物特异性。
巢式PCR能够检测出每1毫升液体中少于10个生物体的DNA。该技术的特异性为96.4%,灵敏度为76.7%。虽然特异性始终很高,但灵敏度取决于所检测的体液:脑脊液为100%,尿液为100%,滑液为80%,血清为59%。假阳性结果率为3.6%。
这些数据证明了PCR在确认莱姆病临床诊断以及深入了解该疾病不同阶段发病机制方面的潜在效用。