Early diagnosis of disseminated adenovirus infection associated with hematopoietic stem cell transplantation by quantitative PCR

定量PCR早期诊断造血干细胞移植相关播散性腺病毒感染

基本信息

批准号：
15591123
负责人：
MORIMOTO Akira
金额：
$ 2.24万
依托单位：
Kyoto Prefectural University of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

A 17-year-old male who had undergone bone marrow transplantation (BMT) died of multiple organ failure. We evaluated his serum and urine samples for adenovirus (AdV) genome using real-time PCR retrospectively. AdV DNA copy numbers, increasing at first in urine followed by in serum, were closely related with his clinical course.A 12-year-old female developed life-threatening pneumonia with hemophagocytic syndrome. We could not pin down pathogen with antibody titer or culture. However, PCR assay of lung tissue was positive for AdV and real-time PCR analysis confirmed this diagnosis.We evaluated serum and urine samples of 7 patients received BMT for AdV genome using real-time PCR in time course retrospectively. With this analysis 3 of 7 patients were confirmed to have disseminated AdV infection. In all of these 3 patients, AdV DNA was found to be positive in urine at one or two weeks after BMT and those were to be more than 10^5 copies/ml in serum at 4 to 6 weeks after BMT. 2 patients, who were found to have more than 10^3 copies/ml AdV in serum before BMT, developed disseminated AdV infection.A 17-year-old female who was received BMT developed intractable diarrhea and spiky fever. Rapid AdV antigen detection test was positive in stool, however, PCR using conventional primers for AdV failed to detect AdV DNA because of 9 base substitutions in this AdV. She was confirmed to have AdV infection by PCR analysis using AdV type 7 specific primers.Quantification of AdV copies is powerful method for early diagnosis of disseminated AdV infection. Patients who are positive for serum AdV copies before BMT and found to be positive for urine AdV copies after BMT are high risk of developing disseminated AdV infection. Using this technique we can detect AdV in biopcied specimens such as lung. However we have to pay attention for false negative results in PCR analysis because AdV has mutations frequently.

一名17岁男性接受骨髓移植（BMT）后死于多器官衰竭。我们使用实时 PCR 技术回顾性评估了他的血清和尿液样本中的腺病毒 (AdV) 基因组。 AdV DNA 拷贝数首先在尿液中增加，随后在血清中增加，这与他的临床病程密切相关。一名 12 岁女性出现了危及生命的肺炎并伴有噬血细胞综合征。我们无法通过抗体滴度或培养物确定病原体。然而，肺组织的 PCR 检测呈 AdV 阳性，实时 PCR 分析证实了这一诊断。我们使用实时 PCR 回顾性评估了 7 名接受 BMT 的患者的血清和尿液样本中的 AdV 基因组。通过此分析，7 名患者中有 3 名被确认患有播散性 AdV 感染。这3例患者均在BMT后1周或2周时尿液中发现AdV DNA呈阳性，且在BMT后4至6周时血清中AdV DNA检测结果超过10^5拷贝/ml。 2例患者在BMT前发现血清中AdV超过10^3拷贝/ml，出现播散性AdV感染。一名17岁女性接受BMT后出现顽固性腹泻和发烧。粪便中AdV抗原快速检测呈阳性，但使用AdV常规引物的PCR未能检测到AdV DNA，因为该AdV中有9个碱基替换。使用AdV 7型特异性引物进行PCR分析，确认她患有AdV感染。AdV拷贝定量是早期诊断播散性AdV感染的有力方法。 BMT 前血清 AdV 拷贝呈阳性且 BMT 后发现尿液 AdV 拷贝呈阳性的患者发生播散性 AdV 感染的风险较高。使用这种技术，我们可以检测肺等活检样本中的 AdV。但由于AdV经常发生突变，因此在PCR分析中必须注意假阴性结果。