Diagnosis of human metapneumovirus by immunofluorescence staining with monoclonal antibodies in the North-East of England
Diagnosis of human metapneumovirus by immunofluorescence staining with monoclonal antibodies in the North-East of England
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DOI：
10.1016/j.jcv.2007.07.018
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发表时间：
2007-11-01
影响因子：
8.8
通讯作者：
Toms, G. L.
中科院分区：
文献类型：
--
作者：
Fenwick, F.;Young, B.;Toms, G. L.
背景：自 2001 年发现以来，人类偏肺病毒 (hMPV) 已被证明是人类呼吸道疾病的重要原因，导致住院儿童呼吸道感染的 5-8%。迄今为止，诊断主要通过逆转录酶聚合酶链反应（RT-PCR）进行，但鼻咽分泌物（IF）细胞的免疫荧光染色为一些实验室提供了优势，并且可以在紧急情况下产生更快速的结果。我们最近证明，IF 与兔抗血清的灵敏度与 RT-PCR 相同。然而，单克隆抗体提供了更丰富、更统一的 IF 试剂。 目的：我们评估了抗 hMPV 单克隆抗体库在住院婴儿和儿童呼吸道感染常规诊断中的应用。 研究设计：使用兔多克隆抗血清和单克隆抗体库并行进行 IF 测试，对 857 例常规呼吸道标本进行了 IF 检测。另外 1003 个样本仅用单克隆抗体池进行了测试。所有标本还通过 IF 检测了一组其他呼吸道病毒。结果：兔多克隆抗血清和单克隆抗体库均在 56 份标本中给出了阳性结果，在 797 份标本中给出了阴性结果。兔多克隆抗体在另外两个样本中检测到病毒，当与单克隆抗体库进行测试时，这两个样本呈阴性，单克隆抗体库的一致性为 96.6%，特异性为 100%。总体而言，5% 的标本中检测到 hMPV，而 18.4% 的标本呈 hRSV 阳性。结论：基于单克隆抗体池的 IF 是一种适合常规使用的稳健检测方法，其灵敏度仅略低于其他可用的主要诊断方法。 (C) 2007 Elsevier B.V. 保留所有权利。
Background: Since its discovery in 2001 human metapneumovirus (hMPV) has been shown to be a significant cause of human respiratory disease, responsible for 5-8% of respiratory infections in hospitalised children. Diagnosis hitherto has been largely carried out by reverse tanscriptase polymerase chain reaction (RT-PCR) but immunofluorescence staining of cells from nasopharyngeal secretions (IF) offers advantages for some laboratories and may produce a more rapid result in urgent cases. We have recently demonstrated that IF with a rabbit antiserum gave sensitivity equal to that of RT-PCR. However, monoclonal antibodies offer a more plentiful, uniform IF reagent.Objectives: Here we have evaluated a pool of anti-hMPV monoclonal antibodies in the routine diagnosis of respiratory infections in hospitalised infants and children.Study design: Eight hundred and fifty-seven routine respiratory specimens were tested by IF with rabbit polyclonal antiserum and monoclonal antibody pool in parallel. A further 1003 specimens were tested with the monoclonal antibody pool alone. All specimens were also tested for a panel of other respiratory viruses by IF.Results: Both rabbit polyclonal antiserum and monoclonal antibody pool gave positive results in 56 and negative results in 797 specimens. The rabbit polyclonal antibody detected virus in a further two specimens which were negative when tested with the monoclonal pool giving a concordance of 96.6% and a specificity of 100% for the monoclonal antibody pool. Overall hMPV was detected in 5% of specimens whilst 18.4% were positive for hRSV.Conclusions: The monoclonal antibody pool-based IF is a robust assay suitable for routine use with a sensitivity only slightly less than that of the other major diagnostic methodologies available. (C) 2007 Elsevier B.V. All rights reserved.