Enhanced detection of infectious airborne influenza virus
Enhanced detection of infectious airborne influenza virus
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DOI：
10.1016/j.jviromet.2011.05.030
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发表时间：
2011-09-01
影响因子：
3.1
通讯作者：
Beezhold, Donald H.
中科院分区：
文献类型：
--
作者：
Blachere, Francoise M.;Cao, Gang;Beezhold, Donald H.
目前用于检测传染性空气传播流感病毒的筛查方法是有限的，并且缺乏灵敏度。为了提高检测气溶胶样品中传染性流感病毒的灵敏度，开发了病毒复制测定法。使用该检测试剂盒，首先通过在Madin-Darby犬肾（MOCK）细胞中复制扩增流感病毒，然后使用定量PCR（qPCR）进行检测。跨越20小时复制期，使用qPCR在几个时间点测量来自感染性病毒的基质基因表达水平，发现呈指数增加。与传统的基于培养的病毒空斑检测相比，病毒复制检测导致流感病毒检测增加4.6 × 10（5）倍。此外，在病毒空斑试验的一半时间内获得病毒复制试验结果。为了证明病毒复制试验能够检测空气传播的流感病毒，将菌株A/WS/33的稀释制剂加载到喷雾器中。在平静空气沉降室内雾化，随后使用NIOSH两级生物气溶胶采样器收集。在对应于2.8E+02/ml的鸡胚感染接近50%终点（CEID（50））的最大稀释浓度下，病毒复制测定能够检测到感染性流感病毒，否则通过病毒空斑测定无法检测到感染性流感病毒。所得结果表明，病毒复制试验在检测气溶胶样品中的传染性流感病毒时具有高度灵敏度。由爱思唯尔公司出版
Current screening methodologies for detecting infectious airborne influenza virus are limited and lack sensitivity. To increase the sensitivity for detecting infectious influenza virus in an aerosol sample, the viral replication assay was developed. With this assay, influenza virus is first amplified by replication in Madin-Darby canine kidney (MOCK) cells followed by detection with quantitative PCR (qPCR). Spanning a 20-h replication period, matrix gene expression levels from infectious virus were measured at several time points using qPCR and found to exponentially increase. Compared with the traditional culture-based viral plaque assay, the viral replication assay resulted in a 4.6 x 10(5) fold increase in influenza virus detection. Furthermore, viral replication assay results were obtained in half the time of the viral plaque assay. To demonstrate that the viral replication assay is capable of detecting airborne influenza virus, dilute preparations of strain A/WS/33 were loaded into a nebulizer. aerosolized within a calm-air settling chamber and subsequently collected using NIOSH Two-Stage Bioaerosol Samplers. At the most diluted concentration corresponding to a chicken embryo infectious close 50% endpoint (CEID(50)) of 2.8E+02/ml, the viral replication assay was able to detect infectious influenza virus that was otherwise undetectable by viral plaque assay. The results obtained demonstrate that the viral replication assay is highly sensitive at detecting infectious influenza virus from aerosol samples. Published by Elsevier B.V.