Upper respiratory tract infections are the world's most common infectious disease. The etiologic agents behind upper respiratory tract infections (URTIs) are, in fact, a diverse set of pathogens such as influenza, parainfluenza, adenovirus, rhinovirus, and others. More than 200 pathogens are known to be involved. Differential diagnosis of viral infections is sometimes complicated by their diversity or similarity of clinical presentation. This work is devoted to the development of a method which enables simultaneous detection of six common viral URTI pathogens: IAV; IBV; RSV; hAdV; hPIV2; and hPIV3. Antibody microarray technology is utilized to accomplish the analysis. In preparation for protein microchip creation, we produced, characterized, and selected approximately 50 monoclonal antibodies; for each of the aforementioned pathogens, an optimal monoclonal antibody pair was selected. A protein microchip was created, and its core working conditions were optimized. With a balance between convenience and maximal assay sensitivity in mind, a one-step analysis approach was developed for accomplishing the ELISA-like "sandwich" interaction on the manufactured microchip (antibody microarray). Reference viral strains were used to establish the lower limits of detection (LoD) for the assay. For IAV, the LoD was 0.25 ng/ml total viral protein. For other viruses, the LoD ranged from 1 to 2 ng/ml total protein. These sensitivity limits are slightly better than those of standard ELISA, but inferior to those of PCR. Overall, we believe that the developed microchip is a good alternative to existing methods, allowing relatively quick (overnight), inexpensive, simultaneous screening of several pathogens. The design of the antibody microarray is conducive to further development, and the panel of analyzed pathogens can be expanded to include approximately 50 members.

中文翻译：

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用于上呼吸道病毒病原体筛查和鉴别诊断的抗体微阵列免疫测定。

上呼吸道感染是世界上最常见的传染病。实际上，上呼吸道感染（URTIs）背后的病原体是多种病原体，例如流感，副流感，腺病毒，鼻病毒等。已知涉及200多种病原体。病毒感染的鉴别诊断有时因其临床表现的多样性或相似性而变得复杂。这项工作致力于开发一种能够同时检测六种常见病毒性URTI病原体的方法：IAV；IBV; RSV；hAdV；hPIV2; 和hPIV3。利用抗体微阵列技术来完成分析。在准备蛋白质微芯片的过程中，我们生产，鉴定并选择了约50种单克隆抗体；对于上述每种病原体，选择了最佳的单克隆抗体对。创建了蛋白质微芯片，并优化了其核心工作条件。考虑到便利性和最大测定灵敏度之间的平衡，开发了一种一步分析方法，用于在制造的微芯片（抗体微阵列）上完成ELISA样的“三明治”相互作用。参考病毒株用于确定检测的下限（LoD）。对于IAV，LoD为0.25 ng / ml总病毒蛋白。对于其他病毒，LoD范围为总蛋白的1到2 ng / ml。这些灵敏度限度比标准ELISA的限度稍好，但不如PCR的限度。总体而言，我们认为开发的微芯片是现有方法的良好替代品，可以相对快速（过夜），价格低廉，同时筛选几种病原体。抗体微阵列的设计有利于进一步的发展，并且所分析病原体的面板可以扩展到包括大约50个成员。