Borg I, Rohde G, Löseke S, Bittscheidt J, Schultze-Werninghaus G, Stephan V, Bufe A
Dept of Experimental Pneumology, Ruhr-University Bochum, and Division of Pneumology, Allergology and Sleep Medicine, University Hospital Bergmannsheil, Bochum, Germany.
Eur Respir J. 2003 Jun;21(6):944-51. doi: 10.1183/09031936.03.00088102.
Respiratory syncytial virus (RSV) is known to cause acute lower respiratory tract infections (ARI) in young children and is involved in exacerbation of chronic obstructive pulmonary disease (COPD) in adults. The role of RSV in stable COPD and the viral load in different respiratory diseases has not been investigated to date. The present authors established and evaluated a quantitative TaqMan real-time polymerase chain reaction assay specific for RSV subgroup A. Absolute quantification for the determination of viral load input was achieved using a control plasmid. The assay allowed for a quantification over a >6-log range and a detection limit of <10 RSV copies per reaction mixture. The assay was 30 times more sensitive than conventional nested polymerase chain reaction assays. Interassay SD was 1.3 and coefficient of variation 4.7% on average. Clinical specimens from infants with ARI (n=62) and elderly adults with COPD (n=125) were compared for viral loads. A total of 47% RSV-positive samples were found in the ARI study group and 28% in the COPD study group. The viral load of both study groups was found to differ significantly. In the ARI study group the viral load was increased almost 2000-fold, suggesting acute infection in this group and former or latent infection in the COPD group. Respiratory syncytial virus-A specific TaqMan real-time polymerase chain reaction assay is a sensitive and rapid method for the determination of viral load in clinical samples. It enables differential statements concerning the involvement of respiratory syncytial virus in acute lower respiratory tract infections and chronic obstructive pulmonary disease to be achieved.
呼吸道合胞病毒（RSV）已知可导致幼儿急性下呼吸道感染（ARI），并与成人慢性阻塞性肺疾病（COPD）的加重有关。迄今为止，RSV在稳定期COPD中的作用以及不同呼吸道疾病中的病毒载量尚未得到研究。本文作者建立并评估了一种针对A亚组RSV的定量TaqMan实时聚合酶链反应检测方法。使用对照质粒实现了对病毒载量输入测定的绝对定量。该检测方法可在>6个对数范围内进行定量，每个反应混合物的检测限<10个RSV拷贝。该检测方法比传统巢式聚合酶链反应检测方法灵敏30倍。批间标准差平均为1.3，变异系数平均为4.7%。比较了ARI婴儿（n=62）和COPD老年成人（n=125）临床标本的病毒载量。ARI研究组共发现47%的RSV阳性样本，COPD研究组为28%。发现两个研究组的病毒载量有显著差异。在ARI研究组中，病毒载量增加了近2000倍，表明该组为急性感染，而COPD组为既往或潜伏感染。呼吸道合胞病毒A特异性TaqMan实时聚合酶链反应检测方法是一种灵敏且快速的临床样本病毒载量测定方法。它能够对呼吸道合胞病毒在急性下呼吸道感染和慢性阻塞性肺疾病中的作用做出不同的判断。
