Noordhoek G T, Wolters E C, de Jonge M E, van Embden J D
Unit Molecular Microbiology, National Institute of Public Health and Environmental Protection, Bilthoven.
J Clin Microbiol. 1991 Sep;29(9):1976-84. doi: 10.1128/jcm.29.9.1976-1984.1991.
A polymerase chain reaction with nested primer pairs based on the DNA sequence of the 39-kDa bmp gene of Treponema pallidum subsp. pallidum is described. The method allowed the detection of purified T. pallidum DNA equivalent to the amount of DNA in a single bacterium and was specific for T. pallidum subspecies. After concentration of DNA, using diatomaceous earth, it was possible to detect about 100 treponemes in 1 ml of cerebrospinal fluid. Cerebrospinal fluid samples from a total of 29 symptomatic and asymptomatic patients with neurosyphilis were tested for the presence of treponemal DNA before and at various intervals after intravenous treatment with penicillin. Prior to the penicillin treatment, we detected T. pallidum DNA in 5 of 7 patients with acute symptomatic neurosyphilis, in none of the 4 patients with chronic symptomatic neurosyphilis tested before treatment, and in 2 of 16 patients with asymptomatic neurosyphilis. Unexpectedly, T. pallidum DNA was also often detected in cerebrospinal fluid long after intervenous treatment with penicillin, sometimes up to 3 years after therapy.
描述了一种基于梅毒螺旋体苍白亚种39-kDa bmp基因DNA序列的巢式引物对聚合酶链反应。该方法能够检测到相当于单个细菌中DNA量的纯化梅毒螺旋体DNA,并且对梅毒螺旋体亚种具有特异性。使用硅藻土浓缩DNA后,有可能在1毫升脑脊液中检测到约100个梅毒螺旋体。对总共29例有症状和无症状神经梅毒患者的脑脊液样本在静脉注射青霉素治疗前及治疗后的不同时间间隔检测梅毒螺旋体DNA的存在情况。在青霉素治疗前,我们在7例急性有症状神经梅毒患者中的5例、治疗前检测的4例慢性有症状神经梅毒患者中均未检测到、以及16例无症状神经梅毒患者中的2例中检测到梅毒螺旋体DNA。出乎意料的是,在静脉注射青霉素治疗后很长时间,有时在治疗后长达3年,脑脊液中也经常检测到梅毒螺旋体DNA。