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用于快速诊断呼吸道腺病毒感染的免疫层析试验原型的开发。

Development of a prototype immunochromatographic test for rapid diagnosis of respiratory adenovirus infection.

作者信息

Paulini Inarei, Siqueira-Silva Joselma, Thomaz Luciana, Rocha Leticia, Harsi Charlotte, Bellei Nancy, Granato Celso

机构信息

Universidade Federal de São Paulo, Departamento de Medicina, Laboratório de Virologia, Disciplina de Infectologia, São Paulo, SP, Brazil.

Universidade de São Paulo, Instituto de Biociências, Departamento de Microbiologia, Laboratório de Adenovírus, São Paulo, SP, Brazil.

出版信息

Braz J Infect Dis. 2017 Sep-Oct;21(5):500-506. doi: 10.1016/j.bjid.2017.03.023. Epub 2017 Jun 15.

Abstract

Human adenoviruses comprise an important group of etiologic agents that are responsible for various diseases in adults and children, such as respiratory, ocular, gastroenteric, and urinary infections. In immunocompromised and organ-transplanted individuals, these agents can cause generalized infections. Rapid diagnostic methods for detecting these infectious agents are not widely available. The aim of this work was to produce monoclonal and polyclonal anti-adenovirus antibodies to be used in a rapid diagnostic test for respiratory infections. Adenovirus hexons were satisfactorily purified by ultracentrifugation and chromatography. After virus purification, anti-hexon monoclonal antibodies were produced and characterized, following classical methods. Antibodies were specific for adenoviruses 2, 3, 5, and 41. The proposed immunochromatographic test was standardized using colloidal gold. The standardization of the rapid test was sufficient to detect adenovirus antigens (in nasopharyngeal lavage samples) with sensitivity of 100% and specificity of 85% when compared to direct immunofluorescence. The immunochromatographic assay prototype was sufficiently sensitive to detect B (3), C (2 and 5), and F (41) adenovirus samples. Although based on preliminary data, the test demonstrated the same performance as direct immunofluorescence, but with the advantage of being a point-of-care test. Further studies are still needed to confirm its effectiveness in clinical practice.

摘要

人腺病毒是一类重要的病原体,可导致成人和儿童的多种疾病,如呼吸道、眼部、胃肠道和泌尿系统感染。在免疫功能低下和器官移植个体中,这些病原体可引起全身性感染。目前广泛可用的检测这些感染因子的快速诊断方法并不多。这项工作的目的是制备单克隆和多克隆抗腺病毒抗体,用于呼吸道感染的快速诊断测试。通过超速离心和色谱法成功纯化了腺病毒六邻体。病毒纯化后,按照经典方法制备并鉴定了抗六邻体单克隆抗体。这些抗体对腺病毒2、3、5和41具有特异性。使用胶体金对所提出的免疫层析试验进行了标准化。与直接免疫荧光法相比,该快速检测方法的标准化足以检测腺病毒抗原(鼻咽灌洗样本中),灵敏度为100%,特异性为85%。免疫层析试验原型对检测B(3型)、C(2型和5型)和F(41型)腺病毒样本具有足够的敏感性。尽管基于初步数据,但该检测方法显示出与直接免疫荧光法相同的性能,但具有即时检测的优势。仍需要进一步研究以证实其在临床实践中的有效性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a3fc/9425546/d87a80ab2e30/gr1.jpg

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