Rapid and Highly Sensitive Method for Influenza A (H1N1) Virus Detection
Rapid and Highly Sensitive Method for Influenza A (H1N1) Virus Detection
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DOI：
10.1021/ac3002947
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发表时间：
2012-05-01
影响因子：
7.4
通讯作者：
Chou, Chien
中科院分区：
文献类型：
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作者：
Su, Li-Chen;Chang, Chung-Ming;Chou, Chien
在这项研究中，我们应用所开发的配对表面等离子体波生物传感器（PSPWB）的双通道生物传感器的快速和灵敏的检测猪源甲型H1N1流感病毒（S-OIV）。结合信号和参考通道的幅度比，PSPWB系统的稳定性显着提高实验。双通道PSPWB检测S-OIV的理论检测限（LOD）为30 PFU/mL（PFU，空斑形成单位），其根据表面等离子体共振信号与磷酸盐缓冲盐水（PBS）中S-OIV临床分离株浓度（18-1.8 x 10（6）PFU/mL）的拟合曲线计算得出。在最差情况下，LOD的灵敏度比市售流感快速诊断检测试剂高2个数量级，比实时定量聚合酶链反应（PCR）低1个数量级，在本实验中，PBS中S-OIV的LOD确定为3.5 PFU/mL。此外，在体内条件下，该实验证明，该测定法成功测量了模拟溶液中浓度为1.8 x 10（2）PFU/mL的S-OIV，该模拟溶液含有PBS稀释的正常人鼻粘膜。最重要的是，检测时间不到20分钟。从结果来看，双通道PSPWB可能提供了一个很大的机会，在开发一种替代的PCR-免费的诊断方法，快速，灵敏，准确地检测病毒病原体与流行病学相关的临床样本，通过使用适当的病原体特异性抗体。
In this study, we applied the developed paired surface plasma waves biosensor (PSPWB) in a dual-channel biosensor for rapid and sensitive detection of swine-origin influenza A (H1N1) virus (S-OIV). In conjunction with the amplitude ratio of the signal and the reference channel, the stability of the PSPWB system is significantly improved experimentally. The theoretical limit of detection (LOD) of the dual-channel PSPWB for S-OIV is 30 PFU/mL (PFU, plaque-forming unit), which was calculated from the fitting curve of the surface plasmon resonance signal with a S-OIV clinical isolate concentration in phosphate-buffered saline (PBS) over a range of 18-1.8 x 10(6) PFU/mL. The LOD is 2 orders of magnitude more sensitive than the commercial rapid influenza diagnostic test at worst and an order of magnitude less sensitive than real-time quantitative polymerase chain reaction (PCR) whose LOD for S-OIV in PBS was determined to be 3.5 PFU/mL in this experiment. Furthermore, under in vivo conditions, this experiment demonstrates that the assay successfully measured S-OIV at a concentration of 1.8 x 10(2) PFU/mL in mimic solution, which contained PBS-diluted normal human nasal mucosa. Most importantly, the assay time took less than 20 min. From the results, the dual-channel PSPWB potentially offers great opportunity in developing an alternative PCR-free diagnostic method for rapid, sensitive, and accurate detection of viral pathogens with epidemiological relevance in clinical samples by using an appropriate pathogen-specific antibody.