Comparison of immunofluorescence with monoclonal antibodies and RT-PCR for the detection of human coronaviruses 229E and OC43 in cell culture
Comparison of immunofluorescence with monoclonal antibodies and RT-PCR for the detection of human coronaviruses 229E and OC43 in cell culture
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DOI：
10.1016/s0166-0934(98)00013-5
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发表时间：
1998-06-01
影响因子：
3.1
通讯作者：
Talbot, PJ
中科院分区：
文献类型：
--
作者：
Sizun, J;Arbour, N;Talbot, PJ
人类冠状病毒与两个已知的血清群229E和OC43一起，导致多达三分之一的普通感冒，并可能与医院内呼吸道感染、小肠结肠炎、心包炎和神经疾病等严重疾病有关。为了更好地了解它们在病理学中的作用，需要在临床样本中采用可靠的检测方法。作为设计这种诊断程序的第一步，在一个实验细胞培养模型中比较了两种病毒诊断方法的敏感性和特异性：使用单抗的间接免疫荧光分析和从感染细胞中提取病毒RNA的逆转录聚合酶链式反应扩增。免疫荧光法在MOI低至10(-2)的细胞中检测到人类冠状病毒(logTCID50/ml分别为4.25和2.0；对数PFU/ml分别为4.83和1.84)，而逆转录聚合酶链式反应扩增为10(-3)(丙型肝炎病毒-OC43)或10(-4)(丙型肝炎病毒)；229E和OC43的对数pfu/ml分别为2.3和1.34)。与其他人类呼吸道病原体：流感病毒、呼吸道合胞病毒和腺病毒没有假阳性信号。此外，每一种检测方法都是冠状病毒血清群特异性的。这些结果表明，用单抗免疫荧光和逆转录聚合酶链式反应RNA扩增快速检测感染细胞培养中的人类冠状病毒具有潜在的实用价值。这两种方法均可用于临床标本的人类感染诊断。(C)1998 Elsevier Science B.V.保留所有权利。
Human coronaviruses, with two known serogroups named 229E and OC43, cause up to one third of common colds and may be associated with serious diseases such as nosocomial respiratory infections, enterocolitis, pericarditis and neurological disorders. Reliable methods of detection in clinical samples are needed for a better understanding of their role in pathology. As a first step in the design of such diagnostic procedures, the sensitivities and specificities of two viral diagnostic assays were compared in an experimental cell culture model: an indirect immune-fluorescence assay using monoclonal antibodies and reverse transcriptase-polymerase chain reaction amplification of viral RNA from infected cells. Immunofluorescence detected human coronaviruses in cells infected at a MOI as low as 10(-2) (log TCID50/ml = 4.25 for HCV-229E and 2.0 for HCV-OC43; log PFU/ml = 4.83 for HCV-229E and 1.84 for HCV-OC43) versus 10(-3) (HCV-OC43) or 10(-4) (HCV-229E) for reverse transcriptase-polymerase chain reaction amplification (log TCID50/ml = 1.75 for HCV-229E and 1.5 for HCV-OC43; log PFU/ml = 2.3 for HCV-229E and 1.34 for HCV-OC43). There were no false positive signals with other human respiratory pathogens: influenza virus, respiratory syncytial virus and adenovirus. Moreover, each assay was coronavirus serogroup-specific. These results demonstrate the potential usefulness of immunofluorescence with monoclonal antibodies and reverse transcriptase-polymerase chain reaction RNA amplification for the rapid detection of human coronaviruses in infected cell cultures. Both methods could be applied to clinical specimens for the diagnosis of human infections. (C) 1998 Elsevier Science B.V. All rights reserved.