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.

一名经历了骨髓移植（BMT）的17岁男性死于多器官衰竭。我们使用实时PCR回顾性地评估了他的血清和尿液样品的腺病毒（ADV）基因组。 ADV DNA拷贝数最初在尿液中增加，随后是血清，与他的临床过程密切相关。一名12岁的女性患有造成血糖综合征的肺炎。我们无法用抗体滴度或培养物限制病原体。然而，肺组织的PCR分析对ADV的阳性为阳性，实时PCR分析证实了该诊断。我们评估了7例患者的血清和尿液样本在回顾性时使用实时PCR接受了ADV基因组的BMT。通过该分析，有7例患者中有3例已传播ADV感染。在所有这3例患者中，在BMT后一两周内发现ADV DNA在尿液中呈阳性，并且在BMT后4至6周时，血清中的ADV DNA应超过10^5份/ml。在BMT之前，有2例患者在血清中有10^3份/ml副词，并发出了散布的ADV感染。受过BMT的17岁雌性患有顽固性的腹泻和尖峰热。快速ADV抗原检测试验在粪便中为阳性，但是，使用常规引物进行ADV的PCR无法检测ADV DNA，因为该ADV中有9个基本取代。她被确认通过使用ADV 7特定引物通过PCR分析具有ADV感染。ADV副本的定量是早期诊断散布ADV感染的有力方法。 BMT之前的血清ADV副本阳性的患者对BMT后的尿液ADV副本呈阳性，这是发展传播ADV感染的高风险。使用此技术，我们可以检测到诸如肺之类的活检标本中的ADV。但是，我们必须注意PCR分析中的虚假负面结果，因为ADV经常具有突变。