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[通过间接免疫荧光法进行爱泼斯坦-巴尔病毒诊断的载玻片研制]

[Development of slides for Epstein-Barr virus diagnosis by indirect immunofluorescence].

作者信息

Pérez Germán R, Taborda Miguel A, Toffi Alicia, Palonsky Marcelo, Pagotto Melina, Gardiol Daniela N, Giri Adriana A

机构信息

Area Virología, Departamento de Microbiología, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario.

出版信息

Medicina (B Aires). 2005;65(4):315-20.

Abstract

Epstein-Barr virus (EBV) is the main oncogenic lymphotropic agent of the Herpesviridae family and is globally distributed. EBV acute infection occurs in young adults producing infectious mononucleosis. Detection of anti-viral capside antigen (VCA) antibodies indicates previous or present EBV infection. Moreover, high titles of anti-VCA antibodies are observed in EBV-associated neoplasic disorders, such as lymphomas in AIDS patients. The objective of this study was the development and optimization of P3HR1 cell slides for the EBV serologic detection by indirect immunofluorescence (IIF) assay. P3HR1 exponential growth culture cells were stimulated with phorbol-12-mirystoil-13-acetate, collected at different time points and used for slide preparation. IIF assay was performed in each slide using an anti-EBV positive serum as primary antibody. An 11% increase in VCA expression was observed at 40 hours post-stimulation. Data was confirmed by Western blot and immunodetection. Intra- and inter-lot precisions of the developed slides were evaluated for IgG and IgM antibodies using EBV-positive sera and positive samples for other members of the Herpesviridae family. Neither false-positive or false negative results were obtained for EBV detection nor cross-reaction was observed with other members of the Herpesviridae family with the developed slides. In conclusion, the slides here presented can be a useful instrument for acute EBV infection diagnosis and for the serologic detection of IgG anti-VCA antibodies in EBV-associated neoplastic disorders.

摘要

爱泼斯坦-巴尔病毒(EBV)是疱疹病毒科主要的嗜淋巴细胞致癌因子,在全球范围内均有分布。EBV急性感染发生在年轻成年人中,可导致传染性单核细胞增多症。抗病毒衣壳抗原(VCA)抗体的检测表明既往或当前存在EBV感染。此外,在与EBV相关的肿瘤性疾病中,如艾滋病患者的淋巴瘤,可观察到高滴度的抗VCA抗体。本研究的目的是开发和优化用于通过间接免疫荧光(IIF)检测进行EBV血清学检测的P3HR1细胞玻片。用佛波醇-12-肉豆蔻酸酯-13-乙酸酯刺激处于指数生长期的P3HR1培养细胞,在不同时间点收集细胞并用于玻片制备。在每张玻片中使用抗EBV阳性血清作为一抗进行IIF检测。刺激后40小时观察到VCA表达增加了11%。数据通过蛋白质印迹法和免疫检测得到证实。使用EBV阳性血清以及疱疹病毒科其他成员的阳性样本,对所开发玻片针对IgG和IgM抗体的批内和批间精密度进行了评估。所开发的玻片在检测EBV时未获得假阳性或假阴性结果,也未观察到与疱疹病毒科其他成员的交叉反应。总之,本文所展示的玻片可成为诊断EBV急性感染以及检测EBV相关肿瘤性疾病中IgG抗VCA抗体血清学的有用工具。

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