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一种用于定量环介导等温扩增检测食源性病原体的新型CMOS图像传感器系统。

A novel CMOS image sensor system for quantitative loop-mediated isothermal amplification assays to detect food-borne pathogens.

作者信息

Wang Tiantian, Kim Sanghyo, An Jeong Ho

机构信息

Department of Bionanotechnology, Gachon University, San 65, Bokjeong-Dong, Sujeong-Gu, Seongnam-Si, Gyeonggi-Do 461-701, Republic of Korea.

Department of Bionanotechnology, Gachon University, San 65, Bokjeong-Dong, Sujeong-Gu, Seongnam-Si, Gyeonggi-Do 461-701, Republic of Korea; Graduate Gachon Medical Research Institute, Gil Medical Center, Inchon, 405-760, Republic of Korea.

出版信息

J Microbiol Methods. 2017 Feb;133:1-7. doi: 10.1016/j.mimet.2016.12.002. Epub 2016 Dec 10.

Abstract

Loop-mediated isothermal amplification (LAMP) is considered as one of the alternatives to the conventional PCR and it is an inexpensive portable diagnostic system with minimal power consumption. The present work describes the application of LAMP in real-time photon detection and quantitative analysis of nucleic acids integrated with a disposable complementary-metal-oxide semiconductor (CMOS) image sensor. This novel system works as an amplification-coupled detection platform, relying on a CMOS image sensor, with the aid of a computerized circuitry controller for the temperature and light sources. The CMOS image sensor captures the light which is passing through the sensor surface and converts into digital units using an analog-to-digital converter (ADC). This new system monitors the real-time photon variation, caused by the color changes during amplification. Escherichia coli O157 was used as a proof-of-concept target for quantitative analysis, and compared with the results for Staphylococcus aureus and Salmonella enterica to confirm the efficiency of the system. The system detected various DNA concentrations of E. coli O157 in a short time (45min), with a detection limit of 10fg/μL. The low-cost, simple, and compact design, with low power consumption, represents a significant advance in the development of a portable, sensitive, user-friendly, real-time, and quantitative analytic tools for point-of-care diagnosis.

摘要

环介导等温扩增技术(LAMP)被认为是传统聚合酶链反应(PCR)的替代技术之一,它是一种低成本、便携式且功耗极低的诊断系统。本研究描述了LAMP技术在实时光子检测以及与一次性互补金属氧化物半导体(CMOS)图像传感器集成的核酸定量分析中的应用。这个新颖的系统作为一个扩增耦合检测平台,依靠CMOS图像传感器,并借助一个用于温度和光源的计算机化电路控制器来工作。CMOS图像传感器捕获穿过传感器表面的光,并使用模数转换器(ADC)将其转换为数字单元。这个新系统监测由扩增过程中的颜色变化引起的实时光子变化。大肠杆菌O157被用作定量分析的概念验证目标,并与金黄色葡萄球菌和肠炎沙门氏菌的结果进行比较,以确认该系统的效率。该系统在短时间内(45分钟)检测到了不同DNA浓度的大肠杆菌O157,检测限为10fg/μL。其低成本、简单且紧凑的设计以及低功耗,代表了在开发用于即时诊断的便携式、灵敏、用户友好、实时和定量分析工具方面的重大进展。

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