Viral diagnosis by antigen detection techniques
Viral diagnosis by antigen detection techniques
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DOI：
10.1016/0928-0197(96)00209-7
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发表时间：
1996-05-01
期刊：
影响因子：
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通讯作者：
Grandien, M
中科院分区：
文献类型：
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作者：
Grandien, M
背景：通过直接在临床标本中检测病毒抗原，可以在疾病的早期阶段获得病毒感染的诊断。方法：用免疫组织化学方法从感染部位采集的细胞中检测病毒感染时产生的抗原，用固相免疫分析法(IA)检测分泌物和血液中的抗原。可在呼吸道细胞中诊断引起急性呼吸道感染的病毒，可通过免疫荧光(IF)或免疫过氧化物酶技术在外周血白细胞(PBL)中检测到引起皮肤刮片上皮细胞水疱疹的病毒、脑神经细胞或皮肤和角膜上皮细胞中的狂犬病病毒以及巨细胞病毒(CMV)基质抗原pp65。在阅读结果的过程中，可以很容易地检查样本的质量。一些用于抗原检测的IAS，如血液中HBs Ag和HIV p24抗原的检测，是标准化和灵敏的。其他人给出的结果不那么敏感，因为临床标本的质量不同。乳胶凝集试验主要用于快速检测粪便中的病毒或病毒抗原：轮状病毒和腺病毒；该方法可能不是很敏感，但在几分钟内就能得出结果。检测病毒核酸的检测比抗原检测更敏感，因为聚合酶链式反应(PCR)获得的基因片段有巨大的扩增。到目前为止，这种检测方法耗时长、费用高，主要用于特定的临床情况。结果：随着特异性单抗的问世，抗原检测技术得到了越来越多的应用。现在，人们认识到需要质量控制、训练有素的工作人员以及标准化的试剂和方法来收集和准备标本。呼吸道病毒在世界各地被用于诊断和流行病学研究。同样，IF仍然是狂犬病诊断最常用的方法。对于CMV，pp65基质抗原被证明是与临床症状密切相关的敏感标志物。用IF技术检测CMV肺炎已被证明优于其他预测骨髓移植患者CMV肺炎的技术。免疫吸附试验目前被用于基于血清样本中抗原检测的大规模诊断的全自动化系统中。利用针对临床标本中最丰富的病毒抗原的单抗提高了固相抗体的特异性，缩短了反应时间；这一点已被大多数不断出现的基于免疫分析原理的新的快速诊断试剂盒所采用。结论：虽然在病毒学中，基因检测方法获得了更敏感的结果，但直接在临床标本中进行的病毒抗原检测试验，在引入单抗用于诊断后，由于对实验室设备的要求低、快速和早期的结果以及相对较低的铸型，被越来越多地使用。抗原检测被成功地直接用于临床标本，用于许多病毒感染的快速诊断以及组织培养分离病毒的鉴定。有了基于单抗的IAS，反应时间缩短了，市场上出现了新的快速、几乎是“即时”的检测试剂盒。
Background: Diagnosis of viral infections can be obtained in the early stages of a disease by detection of viral antigens directly in the clinical specimen. This has become an important tool for rapid virus diagnosis.Methods: Antigens produced during virus infections can be detected either in cells collected from the site of infection by immunohistological investigation or in secretions and blood by solid phase immunoassays (IA). Viruses causing acute respiratory infections can be diagnosed in cells from the respiratory tract, viruses causing vesicular eruptions in epithelial cells from skin scrapings, rabies virus in nerve cells of the brain or epithelial cells from skin and cornea and cytomegalovirus (CMV) matrix antigen, pp65, can be detected in peripheral blood leukocytes (PBL) by immunofluorescence (IF) or immunoperoxidase techniques. The quality of specimens can be easily checked during the reading of results. Some IAs for antigen detection, such as detection of HBsAg and HIV p24 antigen in blood are standardized and sensitive. Others give less sensitive results because of the variation of quality of the clinical specimen. The latex agglutination tests are mainly used for rapid detection of virus or viral antigens in faeces: rota- and adenoviruses; the method may not be very sensitive but yields a result within a few minutes. Assays detecting viral nucleic acids are more sensitive than antigen detection tests because of a tremendous amplification of gene segments obtained by the polymerase chain reaction (PCR). So far such assays are time consuming and expensive and are mainly used in specific clinical situations.Results: After introduction of specific monoclonal antibodies (Mabs), the antigen detection techniques are increasingly used. The need for quality control, trained staff, and standardized reagents and methods for specimen collection and preparation is now being appreciated. IF for viral respiratory viruses is used for diagnosis and epidemiological studies all over the world. Likewise, IF is still the method most often used for rabies diagnosis. For CMV, the pp65 matrix antigen is shown to be a sensitive marker closely correlated with clinical symptoms. Its detection by the IF technique has proven to be superior to other techniques for prediction of CMV pneumonia in bone marrow transplant patients. IAs are currently used in fully automated systems for large scale diagnosis based on antigen detection in serum specimens. Increase of antibody specificity on the solid phase by use of Mabs directed against the most abundant viral antigen in the clinical specimen shortens the reaction time; this has been employed in most of the constantly appearing new rapid diagnosis kits based on the immunoassay principle.Conclusion: Although, in virology, more sensitive results are obtained by the gene detection method, PCR-directly in clinical samples, viral antigen detection tests are, after the introduction of Mabs for diagnostic purposes, increasingly used because of their low demand on laboratory equipment, their rapid and early result and relatively low cast. Antigen detection is successfully used directly in clinical specimens for rapid diagnosis of many viral infections as well as for identification of tissue culture isolated viruses. With Mab-based IAs the reaction time is shortened and new rapid, almost 'instant test' kits are appearing on the market.