Detection & Viral Load Monitoring of Adenovirus

第一阶段项目描述(由申请人提供)：腺病毒(ADV)是越来越多的人认识到的病毒病原体，可导致显著的发病率和死亡率，特别是在免疫功能低下的患者中。据报道，免疫缺陷患者的肺炎死亡率高达60%，肝炎高达50%，总体死亡率高达25%。对这些患者的严重ADV感染的处理需要早期诊断，与其他情况相鉴别，以及在治疗后监测感染的能力。不幸的是，目前还没有针对ADV的快速、准确和定量的诊断技术。ViraCor希望为临床医生提供一种新的诊断工具，它将彻底改变ADV感染的诊断和管理方式，从而降低死亡率。该项目的目的是揭示定量、实时聚合酶链式反应技术是否可以应用于ADV，以产生一种能够提供准确和早期检测病毒的诊断测试。 申请人的初步工作表明，针对所有ADV共享的ADV基因组六邻体区域的引物将对病毒的分子检测具有高度的敏感性和特异性。待研究完成后，如未能检测到所有ADV毒株，调查人员便会进行样本类型、样本处理和DNA提取方法的优化。在这些方面存在潜在的显著差异，这可能会改变患者对总病毒载量的总体印象。同时，申请者将确定分析的性能特征：范围、线性、重现性和测定内和测定间条件的变异系数。达到这些目标后，研究人员将前往他们的研究合作者那里，从培养证实的ADV感染患者那里获得回顾性标本，以开始将定量实验室数据与临床表现联系起来。其目标是显示优化样本的总病毒载量与临床画面的严重程度和进展相关。

DESCRIPTION OF PHASE I PROJECT (provided by applicant): Adenoviruses (AdVs) are increasingly recognized as viral pathogens that can cause significant morbidity and mortality, especially in the immunocompromised patient. Mortality rates in the immunocompromised patient have been reported to be as high as 60 percent for pneumonia, 50 percent for hepatitis and 25 percent overall. Management of serious AdV infections in these patients requires early diagnosis, differentiation from other conditions and the ability to monitor the infection after treatment. Unfortunately, rapid, accurate and quantitative diagnostic techniques for AdV do not exist at this time. ViraCor would like to offer the clinician a new diagnostic tool that will revolutionize the way AdV infections are diagnosed and managed, thereby reducing mortality rates. The aim of the project is to uncover whether quantitative, real-time PCR technology can be applied to AdV to produce a diagnostic test that can provide accurate and early detection of the virus. The applicants' preliminary work indicates that primers against the hexon region of the AdV genome, which is shared by all the AdVs, will be highly sensitive and specific for the molecular detection of the virus. Upon completion of the studies showing that the primers can detect most, if not all the AdV strains, the investigators will proceed to optimization of specimen types, specimen processing and DNA isolation methods. There are potentially significant differences in these areas, which could change the overall impression of total viral load in the patient. Concurrently, the applicants will be establishing performance characteristics of the assay: range, linearity, reproducibility, and coefficient of variation for intra-assay and inter-assay conditions. After meeting these goals the investigators will go to their study collaborators and obtain retrospective specimens from patients with culture proven AdV infections to begin correlation of quantitative laboratory data with clinical presentation. The goal is to show total viral load from optimized specimens correlates with severity and progression of the clinical picture.