MOLECULAR STRATEGIES FOR CONGENITAL SYPHILIS DIAGNOSIS

这项建议主要目标是(1)重新完善 发展了分子方法学(IgM免疫印迹、聚合酶链法 聚合酶链式反应在先天梅毒诊断和检测中的应用 梅毒螺旋体入侵中枢神经系统，(2)利用 基因工程克隆梅毒螺旋体膜免疫原AS 酶联免疫吸附试验(ELISA)检测的基础 新生儿抗T。梅毒IgM抗体，以及(3)测定梅毒抗体 在临床环境中，这些化验方法将具有最大的 对病人管理的影响。这项研究将集中在 梅毒螺旋体症状性感染婴儿的鉴定； 初步研究支持这样一种观点，即与 有症状的婴儿，没有临床或实验室参数 目前正在使用，可以推断为无症状的婴儿 确认为先天性感染。因此，对症状和症状的评估 无症状的婴儿需要完全不同的诊断 接近了。 未经治疗或治疗不当的梅毒母亲所生的婴儿 将对血清和脑脊液进行特异性IgM检测 IgM免疫印迹和IgM-EL ISA以及特异性抗体 用聚合酶链式反应检测梅毒螺旋体DNA。这些研究的结果将是 通过平行实验证实和验证了 给兔子接种以确认存在活的梅毒螺旋体I 临床标本。在这种情况下，灵敏度和特异度 可建立IgM免疫印迹法、IgM-ELISA法和聚合酶链式反应。我们计划 改进这些方法和途径，以便更全面地进行评估 聚合酶链式反应与免疫球蛋白M联合检测的正确应用 免疫印迹和IgM-ELISA法诊断先天性梅毒 高危婴儿的特定亚群。改进的诊断 最终将提供更多的发病机制和流行病学数据，所有 这将导致对病例定义、患者策略的改进 管理、治疗选择和后续管理。

The principle objective of this proposal are (1) to refine newly developed molecular methodologies (IgM immunoblotting, polymerase chain reaction [PCR] for the diagnosis of congenital syphilis and detection of central nervous system invasion by Treponema pallidum, (2) to utilize genetically-engineered forms of clone T. pallidum membrane immunogens as the basis of an enzyme-linked immunosorbent assay (ELISA) for detection of neonatal anti-T. pallidum IgM antibodies, and (3) to determine the clinical circumstances in which these assays will have the greatest impact on patient management. This study will focus on the identification of infants symptomatically infect with T. pallidum; preliminary studies have supported the contention that, in contrast to the symptomatic infant, there is no clinical or laboratory parameter currently in use which can be extrapolated to the asymptomatic infant to confirm congenital infection. Therefore, evaluation of symptomatic and asymptomatic infants will require fundamentally different diagnostic approaches. Infants born to mother with untreated or inadequately treated syphilis will have serum and cerebrospinal fluid tested for specific IgM antibodies by IgM immunoblotting and IgM ELISA, as well as for specific T. pallidum DNA by PCR. The results of these studies will be corroborated and validated through parallel experiments involving the inoculation of rabbits to confirm the presence of viable T. pallidum i the clinical specimen. In this way, the sensitivity and specificity of IgM immunoblotting, IgM ELISA, and PCR can be established. We plan to refine these methods and approaches in order to evaluated more completely the appropriate application of the PCR in combination with IgM immunoblotting and IgM ELISA for the diagnosis of congenital syphilis in specific subpopulations of at-risk infants. Improved diagnostics ultimately will provide more pathogenesis and epidemiologic data, all of which will lead to refinements in strategies for case definition, patient management, treatment options, and follow-up management.