Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, VIC 3220, Australia.
Biomed Res Int. 2013;2013:289295. doi: 10.1155/2013/289295. Epub 2013 Feb 6.
Microsphere suspension array systems enable the simultaneous fluorescent identification of multiple separate nucleotide targets in a single reaction. We have utilized commercially available oligo-tagged microspheres (Luminex MagPlex-TAG) to construct and evaluate multiplexed assays for the detection and differentiation of Hendra virus (HeV) and Nipah virus (NiV). Both these agents are bat-borne zoonotic paramyxoviruses of increasing concern for veterinary and human health. Assays were developed targeting multiple sites within the nucleoprotein (N) and phosphoprotein (P) encoding genes. The relative specificities and sensitivities of the assays were determined using reference isolates of each virus type, samples from experimentally infected horses, and archival veterinary diagnostic submissions. Results were assessed in direct comparison with an established qPCR. The microsphere array assays achieved unequivocal differentiation of HeV and NiV and the sensitivity of HeV detection was comparable to qPCR, indicating high analytical and diagnostic specificity and sensitivity.
微球悬浮阵列系统能够在单个反应中同时荧光识别多个单独的核苷酸靶标。我们已经利用市售的寡核苷酸标记微球（Luminex MagPlex-TAG）构建并评估了用于检测和区分亨德拉病毒（HeV）和尼帕病毒（NiV）的多重分析。这两种病原体都是蝙蝠传播的人畜共患病副粘病毒，对兽医和人类健康的关注日益增加。针对核蛋白（N）和磷蛋白（P）编码基因中的多个位点开发了分析。使用每种病毒类型的参考分离株、来自实验感染马的样本和存档兽医诊断提交物来确定分析的相对特异性和敏感性。结果与已建立的 qPCR 直接比较进行评估。微球阵列分析能够明确地区分 HeV 和 NiV，并且 HeV 的检测灵敏度与 qPCR 相当，表明具有高分析和诊断特异性和敏感性。
