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基于局域表面等离子体耦合荧光光纤生物传感器检测猪源流感 A(H1N1)病毒。

Detection of swine-origin influenza A (H1N1) viruses using a localized surface plasmon coupled fluorescence fiber-optic biosensor.

机构信息

Institute of Biophotonics, National Yang-Ming University, Taipei 112, Taiwan.

出版信息

Biosens Bioelectron. 2010 Nov 15;26(3):1068-73. doi: 10.1016/j.bios.2010.08.060. Epub 2010 Aug 26.

Abstract

Swine-origin influenza A (H1N1) virus (S-OIV) was identified as a new reassortant strain of influenza A virus in April 2009 and led to an influenza pandemic. Accurate and timely diagnoses are crucial for the control of influenza disease. We developed a localized surface plasmon coupled fluorescence fiber-optic biosensor (LSPCF-FOB) which combines a sandwich immunoassay with the LSP technique using antibodies against the hemagglutinin (HA) proteins of S-OIVs. The detection limit of the LSPCF-FOB for recombinant S-OIV H1 protein detection was estimated at 13.9 pg/mL, which is 10(3)-fold better than that of conventional capture ELISA when using the same capture antibodies. For clinical S-OIV isolates measurement, meanwhile, the detection limit of the LSPCF-FOB platform was calculated to be 8.25 × 10(4)copies/mL, compared with 2.06 × 10(6)copies/mL using conventional capture ELISA. Furthermore, in comparison with the influenza A/B rapid test, the detection limit of the LSPCF-FOB for S-OIV was almost 50-fold in PBS solution and 25-fold lower in mimic solution, which used nasal mucosa from healthy donors as the diluent. The findings of this study therefore indicate that the high detection sensitivity and specificity of the LSPCF-FOB make it a potentially effective diagnostic tool for clinical S-OIV infection and this technique has the potential to be applied to the development of other clinical microbe detection platforms.

摘要

猪源甲型流感病毒(S-OIV)于 2009 年 4 月被鉴定为一种新型甲型流感病毒重组株,并引发了流感大流行。准确、及时的诊断对于控制流感疾病至关重要。我们开发了一种局部表面等离子体耦合荧光光纤生物传感器(LSPCF-FOB),该传感器将夹心免疫测定法与 LSP 技术相结合,使用针对 S-OIV 的血凝素(HA)蛋白的抗体。LSPCF-FOB 对重组 S-OIV H1 蛋白检测的检测限估计为 13.9pg/mL,当使用相同的捕获抗体时,比传统捕获 ELISA 好 10(3)倍。对于临床 S-OIV 分离物的测量,同时,LSPCF-FOB 平台的检测限计算为 8.25×10(4)拷贝/mL,而传统捕获 ELISA 的检测限为 2.06×10(6)拷贝/mL。此外,与流感 A/B 快速检测相比,LSPCF-FOB 对 S-OIV 的检测限在 PBS 溶液中几乎高 50 倍,在模拟溶液中低 25 倍,模拟溶液使用健康供体的鼻黏膜作为稀释剂。因此,本研究的结果表明,LSPCF-FOB 的高检测灵敏度和特异性使其成为临床 S-OIV 感染的潜在有效诊断工具,并且该技术有可能应用于其他临床微生物检测平台的开发。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d065/7126842/f94decdff179/gr1.jpg

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