Andresen Dennie, von Nickisch-Rosenegk Markus, Bier Frank F
Fraunhofer Institute for Biomedical Engineering (IBMT), Branch Potsdam, Department of Nanobiotechnology and Nanomedicine, Am Muehlenberg 13, 14476 Potsdam-Golm, Germany.
Clin Chim Acta. 2009 May;403(1-2):244-8. doi: 10.1016/j.cca.2009.03.021. Epub 2009 Mar 18.
The need for fast, specific and sensitive multiparametric detection methods is an ever growing demand in molecular diagnostics. Here we report on a newly developed method, the helicase dependent OnChip amplification (OnChip-HDA). This approach integrates the analysis and detection in one single reaction thus leading to time and cost savings in multiparametric analysis.
HDA is an isothermal amplification method that is not depending on thermocycling as known from PCR due to the helicases' ability to unwind DNA double-strands. We have combined the HDA with microarray based detection, making it suitable for multiplex detection. As an example we used the OnChip HDA in single and multiplex amplifications for the detection of the two pathogens N. gonorrhoeae and S. aureus directly on surface bound primers.
We have successfully shown the OnChip-HDA and applied it for single- and duplex-detection of the pathogens N. gonorrhoeae and S. aureus.
We have developed a new method, the OnChip-HDA for the multiplex detection of pathogens. Its simplicity in reaction setup and potential for miniaturization and multiparametric analysis is advantageous for the integration in miniaturized Lab on Chip systems, e.g. needed in point of care diagnostics.
在分子诊断领域，对快速、特异且灵敏的多参数检测方法的需求不断增长。在此，我们报告一种新开发的方法——解旋酶依赖性芯片上扩增（OnChip-HDA）。这种方法将分析与检测整合于一个单一反应中，从而在多参数分析中节省时间和成本。
HDA是一种等温扩增方法，由于解旋酶具有解开DNA双链的能力，它不像PCR那样依赖热循环。我们将HDA与基于微阵列的检测相结合，使其适用于多重检测。例如，我们使用芯片上HDA进行单重和多重扩增，直接在表面结合引物上检测两种病原体——淋病奈瑟菌和金黄色葡萄球菌。
我们成功展示了芯片上HDA，并将其应用于淋病奈瑟菌和金黄色葡萄球菌病原体的单重和双重检测。
我们开发了一种用于病原体多重检测的新方法——芯片上HDA。其反应设置简单，具有小型化和多参数分析的潜力，有利于集成到小型化的芯片实验室系统中，例如即时诊断所需的系统。
