MOLECULAR STRATEGIES FOR CONGENITAL SYPHILIS DIAGNOSIS

本提案的主要目标是（1）重新完善 开发了分子方法学（IgM免疫印迹、聚合酶链 先天性梅毒的诊断和检测 梅毒螺旋体侵入中枢神经系统，（2）利用 克隆T的基因工程形式。苍白球膜免疫原 酶联免疫吸附试验（ELISA）检测的基础 新生儿抗T抗体梅毒IgM抗体，和（3）以确定 这些测定将具有最大的临床环境 对患者管理的影响。 本研究将集中于 婴幼儿弓形虫感染鉴定苍白球; 初步研究支持了这样的论点，即与 有症状的婴儿，没有临床或实验室参数 目前使用的方法可以外推到无症状的婴儿， 确认是先天性感染 因此，评估症状和 无症状的婴儿将需要根本不同的诊断 接近。 母亲患有未经治疗或治疗不当的梅毒所生婴儿 将对血清和脑脊液进行特异性IgM检测 IgM免疫印迹和IgM ELISA抗体，以及特异性 T. PCR检测苍白球DNA。 这些研究的结果将是 通过平行实验证实和验证， 接种家兔以确认活T.苍白球Ⅰ 临床标本。 通过这种方式， 可建立IgM免疫印迹、IgM ELISA和PCR。 我们计划 完善这些方法和途径，以便更全面地评估 PCR联合IgM的适当应用 免疫印迹法和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， 这将导致对病例定义、患者 管理、治疗选择和随访管理。

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.