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在英格兰东北部采用单克隆抗体免疫荧光染色法诊断人偏肺病毒

Diagnosis of human metapneumovirus by immunofluorescence staining with monoclonal antibodies in the North-East of England.

作者信息

Fenwick F, Young B, McGuckin R, Robinson M J, Taha Y, Taylor C E, Toms G L

机构信息

The School of Clinical Medical Sciences, The University of Newcastle upon Tyne, Newcastle upon Tyne NE2 4HH, United Kingdom.

出版信息

J Clin Virol. 2007 Nov;40(3):193-6. doi: 10.1016/j.jcv.2007.07.018. Epub 2007 Sep 14.

Abstract

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.

摘要

背景

自2001年被发现以来,人偏肺病毒(hMPV)已被证明是人类呼吸道疾病的一个重要病因,在住院儿童的呼吸道感染中占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,但对鼻咽分泌物细胞进行免疫荧光染色(IF)对一些实验室来说具有优势,并且在紧急情况下可能会得出更快的结果。我们最近证明,用兔抗血清进行免疫荧光染色的敏感性与RT-PCR相当。然而,单克隆抗体提供了一种更丰富、更均匀的免疫荧光试剂。

目的

在此,我们评估了一组抗hMPV单克隆抗体在住院婴幼儿和儿童呼吸道感染常规诊断中的应用。

研究设计

857份常规呼吸道标本同时用兔多克隆抗血清和单克隆抗体组合进行免疫荧光检测。另外1003份标本仅用单克隆抗体组合进行检测。所有标本还通过免疫荧光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组合在56份标本中呈阳性结果,在797份标本中呈阴性结果。兔多克隆抗体在另外两份标本中检测到病毒,而用单克隆抗体组合检测时这两份标本为阴性,单克隆抗体组合的一致性为96.6%,特异性为100%。总体而言,5%的标本检测到hMPV,而18.4%的标本hRSV呈阳性。

结论

基于单克隆抗体组合的免疫荧光检测是一种可靠的检测方法,适合常规使用,其敏感性仅略低于其他主要可用诊断方法。

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