Efficient, specific, compact hepatitis B diagnostic device: Optical detection of the hepatitis B virus by isothermal amplification
Efficient, specific, compact hepatitis B diagnostic device: Optical detection of the hepatitis B virus by isothermal amplification
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DOI：
10.1016/j.snb.2007.05.015
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发表时间：
2007-11-15
影响因子：
8.4
通讯作者：
Lin, Chii-Wann
中科院分区：
文献类型：
--
作者：
Lee, Szu-Yuan;Lee, Chun-Nan;Lin, Chii-Wann
B型肝炎病毒（HBV）是一种常见的病毒病原体，感染全世界超过3.7亿人。已证实HBV感染与从慢性肝炎到肝硬化或肝细胞癌的其他临床问题相关。一种能够快速准确诊断传染病的微型生物传感器将是无价的，需求量很大。在这项研究中，我们证明了环介导的等温扩增（LAMP）反应，等温核酸扩增方法，在25 μ l的微型反应器中的HBV病毒DNA的扩增和光学检测的可行性。微反应器由多层聚甲基丙烯酸甲酯（PMMA）制成，并且其包括外部温度控制器和产生副产物浊度信号的定量输出的光学检测单元。设计了六种HBV引物和工作缓冲液，并将其预装到反应器中，以使该过程易于操作。实验结果表明，成功的HBV DNA扩增有效（在60分钟），具有高特异性（12种不同的病毒DNA，RNA和2个内部对照）和灵敏度（50拷贝/管）。我们的紧凑型设备是一种集成产品，在包括病原体检测在内的广泛现场应用中具有优势。SNP基因分型和其他基因检测。(c)2007 Elsevier B. V.保留所有权利。
Hepatitis B virus (HBV) is a common viral pathogen that infects over 370 million people worldwide. It has been proven that HBV infection is associated with other clinical problems ranging from chronic hepatitis to cirrhosis or hepatocellular carcinoma. A miniature biosensor that can diagnose infectious diseases rapidly and precisely will be invaluable and in high demand. In this study, we demonstrate the feasibility of a loop-mediated isothermal amplification (LAMP) reaction, an isothermal nucleic acid amplification method, for the amplification and optical detection of HBV viral DNA in a 25 mu l miniature reactor. The micro-reactor is made from multilayer polymethyl methacrylate (PMMA), and it includes an external temperature controller and an optical detection unit that yields quantitative output of a by-product turbidity signal. Six HBV primers and working buffers were designed and are preloaded into the reactor to make the process easy to handle. Experimental results show the successful amplification of HBV DNA efficiently (in 60 min), with high specificity (12 different viral DNA, RNA and 2 internal controls) and sensitivity (50 copies/tube). Our compact device is an integrated product that can be advantageous in a wide spectrum of field applications, including pathogens detection. SNP genotyping and other gene testing. (c) 2007 Elsevier B.V. All rights reserved.