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基于 RT-qPCR 的低密度基因芯片对禽流感病毒进行改良分型：'Riems 流感病毒分型基因芯片 2.0 版'（RITA-2）。

Improved Subtyping of Avian Influenza Viruses Using an RT-qPCR-Based Low Density Array: 'Riems Influenza a Typing Array', Version 2 (RITA-2).

机构信息

Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.

Department of Poultry Diseases, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt.

出版信息

Viruses. 2022 Feb 17;14(2):415. doi: 10.3390/v14020415.

DOI:10.3390/v14020415
PMID:35216008
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8879595/
Abstract

Avian influenza virus (AIV) variants emerge frequently, which challenges rapid diagnosis. Appropriate diagnosis reaching the sub- and pathotype level is the basis of combatting notifiable AIV infections. Real-time RT-PCR (RT-qPCR) has become a standard diagnostic tool. Here, a total of 24 arrayed RT-qPCRs is introduced for full subtyping of 16 hemagglutinin and nine neuraminidase subtypes of AIV. This array, designated Riems Influenza A Typing Array version 2 (RITA-2), represents an updated and economized version of the RITA-1 array previously published by Hoffmann et al. RITA-2 provides improved integration of assays (24 instead of 32 parallel reactions) and reduced assay volume (12.5 µL). The technique also adds RT-qPCRs to detect Newcastle Disease (NDV) and Infectious Bronchitis viruses (IBV). In addition, it maximizes inclusivity (all sequences within one subtype) and exclusivity (no intersubtypic cross-reactions) as shown in validation runs using a panel of 428 AIV reference isolates, 15 reference samples each of NDV and IBV, and 122 clinical samples. The open format of RITA-2 is particularly tailored to subtyping influenza A virus of avian hosts and Eurasian geographic origin. Decoupling and re-arranging selected RT-qPCRs to detect specific AIV variants causing epizootic outbreaks with a temporal and/or geographic restriction is possible.

摘要

禽流感病毒 (AIV) 变体经常出现，这对快速诊断构成了挑战。适当的诊断达到亚型和亚型水平是防治流感病毒感染的基础。实时 RT-PCR（RT-qPCR）已成为一种标准诊断工具。在这里，总共介绍了 24 个排列的 RT-qPCR，用于对 AIV 的 16 种血凝素和 9 种神经氨酸酶亚型进行全部分型。该阵列被命名为 Riems 流感 A 分型阵列版本 2（RITA-2），是 Hoffmann 等人之前发表的 RITA-1 阵列的更新和简化版本。RITA-2 提供了改进的检测集成（24 个而不是 32 个平行反应）和减少的检测体积（12.5 µL）。该技术还增加了用于检测新城疫（NDV）和传染性支气管炎病毒（IBV）的 RT-qPCR。此外，如使用 428 个 AIV 参考分离株、15 个 NDV 和 IBV 参考样本以及 122 个临床样本的验证运行所示，它最大限度地提高了包容性（同一亚型内的所有序列）和排他性（没有亚型间交叉反应）。RITA-2 的开放格式特别适合对禽宿主和欧亚地理起源的甲型流感病毒进行分型。可以对特定的 RT-qPCR 进行解耦和重新排列，以检测具有时间和/或地理限制的导致动物传染病爆发的特定 AIV 变体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/a7ab2c35e21f/viruses-14-00415-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/4bb3b181c640/viruses-14-00415-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/630aef218ca9/viruses-14-00415-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/5ec4572a73a8/viruses-14-00415-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/34e6a5dceffe/viruses-14-00415-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/c551baf71037/viruses-14-00415-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/cb7e7c7b331e/viruses-14-00415-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/4c20272fa2d8/viruses-14-00415-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/a7ab2c35e21f/viruses-14-00415-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/4bb3b181c640/viruses-14-00415-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/630aef218ca9/viruses-14-00415-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/5ec4572a73a8/viruses-14-00415-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/34e6a5dceffe/viruses-14-00415-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/c551baf71037/viruses-14-00415-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/cb7e7c7b331e/viruses-14-00415-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/4c20272fa2d8/viruses-14-00415-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb9d/8879595/a7ab2c35e21f/viruses-14-00415-g008.jpg

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