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

一名17岁的男性接受骨髓移植（BMT）后死于多器官衰竭。我们使用实时PCR回顾性地评估了他的血清和尿液样本中的腺病毒（AdV）基因组。AdV DNA拷贝数在尿液中先升高后升高，与临床病程密切相关。1例12岁女性患者出现危及生命的肺炎伴噬血细胞综合征。我们不能用抗体滴度或培养来确定病原体。然而，肺组织PCR检测阳性AdV和实时PCR分析证实了这一诊断。我们评估了7例患者的血清和尿液样本接受骨髓移植AdV基因组实时PCR在时间过程中回顾性。通过该分析，7例患者中有3例被证实患有播散性AdV感染。3例患者骨髓移植后1 ~ 2周尿中AdV DNA均为阳性，4 ~ 6周血清中AdV DNA均> 10^5拷贝/ml。2例患者骨髓移植前血清AdV含量> 10^3拷贝/ml，发生播散性AdV感染，1例17岁女性患者骨髓移植后出现顽固性腹泻和发热。粪便中AdV抗原快速检测呈阳性，常规AdV引物PCR未能检出AdV DNA，AdV 7型特异性引物PCR检测证实为AdV感染，AdV拷贝数定量是早期诊断播散性AdV感染的有效方法。骨髓移植前血清AdV拷贝阳性，骨髓移植后尿AdV拷贝阳性的患者发生播散性AdV感染的风险较高。使用这种技术，我们可以检测活检标本，如肺腺病毒。但由于腺病毒基因突变频繁，PCR分析中的假阴性结果必须引起重视。

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.