Label-free multiplexed virus detection using spectral reflectance imaging.
Label-free multiplexed virus detection using spectral reflectance imaging.
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DOI：
10.1016/j.bios.2011.01.019
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发表时间：
2011-04-15
影响因子：
12.6
通讯作者：
Unlü MS
中科院分区：
文献类型：
--
作者：
Lopez CA;Daaboul GG;Vedula RS;Ozkumur E;Bergstein DA;Geisbert TW;Fawcett HE;Goldberg BB;Connor JH;Unlü MS
我们展示了一个新的无标记平台的基础上光谱反射成像检测整个病毒和病毒蛋白。干涉反射成像传感器（IRIS）已被证明能够以真实的时间和高通量格式进行灵敏的蛋白质和DNA检测。水泡性口炎病毒（VSV）被用作检测目标，因为它的蛋白质组成特征良好，并且可以被修饰以表达来自其他危险的高致病性因子的病毒外壳蛋白，用于替代检测，同时保持生物安全性2级因子。我们证明了完整的VSV病毒体的特异性检测实现与表面固定的抗体作为捕获探针，这是确认使用荧光成像。确认检测限低至3.5×105空斑形成单位/mL（PFU/mL）。为了增加临床情况下的特异性，外部糖蛋白和内部病毒蛋白质都用相同的抗体阵列用去污剂破坏的纯化VSV和感染的细胞裂解物溶液同时检测。我们的研究结果表明，灵敏和特异性的病毒检测与一个简单的表面化学和最小的样品制备上的定量无标记干涉平台。
We demonstrate detection of whole viruses and viral proteins with a new label-free platform based on spectral reflectance imaging. The Interferometric Reflectance Imaging Sensor (IRIS) has been shown to be capable of sensitive protein and DNA detection in a real time and high-throughput format. Vesicular stomatitis virus (VSV) was used as the target for detection as it is well-characterized for protein composition and can be modified to express viral coat proteins from other dangerous, highly pathogenic agents for surrogate detection while remaining a biosafety level 2 agent. We demonstrate specific detection of intact VSV virions achieved with surface-immobilized antibodies acting as capture probes which is confirmed using fluorescence imaging. The limit of detection is confirmed down to 3.5×105 plaque-forming units/mL (PFUs/mL). To increase specificity in a clinical scenario, both the external glycoprotein and internal viral proteins were simultaneously detected with the same antibody arrays with detergent-disrupted purified VSV and infected cell lysate solutions. Our results show sensitive and specific virus detection with a simple surface chemistry and minimal sample preparation on a quantitative label-free interferometric platform.
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影响因子：
11.8
作者：
Hazelton PR;Gelderblom HR
通讯作者：
Gelderblom HR
影响因子：
2.9
作者：
Burgess, Stewart T. G.;Kenyon, Fiona;Campbell, Colin J.
通讯作者：
Campbell, Colin J.
影响因子：
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作者：
Bisoffi, M.;Hjelle, B.;Larson, R. S.
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Larson, R. S.
影响因子：
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作者：
Boonham, N;Walsh, K;Barker, I
通讯作者：
Barker, I
影响因子：
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作者：
Tani, Hideki;Komoda, Yasumasa;Matsuura, Yoshiharu
通讯作者：
Matsuura, Yoshiharu