Canadian Food Inspection Agency, National Centres for Animal Disease, Lethbridge Laboratory, Township Road 9-1, P.O. Box 640, Lethbridge, Alberta T1J 3Z4, Canada.
J Virol Methods. 2012 Nov;185(2):244-53. doi: 10.1016/j.jviromet.2012.07.005. Epub 2012 Jul 13.
Microarrays are suitable for multiplexed detection and typing of pathogens. Avian influenza virus (AIV) is currently classified into 16 H (hemagglutinin) and 9 N (neuraminidase) subtypes, whereas Newcastle disease virus (NDV) strains differ in virulence and are broadly classified into high and low pathogenicity types. In this study, three assays for detection and typing of poultry viruses were developed on an automated microarray platform: a multiplex assay for simultaneous detection of AIV and detection and pathotyping of NDV, and two separate assays for differentiating all AIV H and N subtypes. The AIV-NDV multiplex assay detected all strains in a 63 virus panel, and accurately typed all high pathogenicity NDV strains tested. A limit of detection of 10(1)-10(3) TCID(50)/mL and 200-400 EID(50)/mL was obtained for NDV and AIV, respectively. The AIV typing assays accurately typed all 41 AIV strains and a limit of detection of 4-200 EID(50)/mL was obtained. Assay validation showed that the microarray assays were generally comparable to real-time RT-PCR. However, the AIV typing microarray assays detected more positive clinical samples than the AIV matrix real-time RT-PCR, and also provided information regarding the subtype. The AIV-NDV multiplex and AIV H typing microarray assays detected mixed infections and could be useful for detection and typing of AIV and NDV.
微阵列适用于病原体的多重检测和分型。禽流感病毒 (AIV) 目前分为 16 种 H（血凝素）和 9 种 N（神经氨酸酶）亚型，而新城疫病毒 (NDV) 株在毒力上存在差异，广泛分为高致病性和低致病性类型。在这项研究中，我们在自动化微阵列平台上开发了三种用于检测和分型家禽病毒的检测方法：一种用于同时检测 AIV 和检测和分型 NDV 的多重检测方法，以及两种用于区分所有 AIV H 和 N 亚型的单独检测方法。AIV-NDV 多重检测方法检测了 63 种病毒面板中的所有毒株，并准确地对所有测试的高致病性 NDV 毒株进行了分型。NDV 和 AIV 的检测限分别为 10(1)-10(3)TCID(50)/mL 和 200-400EID(50)/mL。AIV 分型检测方法准确地对所有 41 株 AIV 毒株进行了分型，检测限为 4-200EID(50)/mL。检测方法验证表明，微阵列检测方法与实时 RT-PCR 通常具有可比性。然而，AIV 分型微阵列检测方法检测到的阳性临床样本比 AIV 基质实时 RT-PCR 多，并且还提供了有关亚型的信息。AIV-NDV 多重和 AIV H 分型微阵列检测方法可检测混合感染，可用于 AIV 和 NDV 的检测和分型。
