Dennie Andresen1, Markus von Nickisch-Rosenegk, Frank F Bier. 1. Fraunhofer Institute for Biomedical Engineering (IBMT), Branch Potsdam, Department of Nanobiotechnology and Nanomedicine, Am Muehlenberg 13, 14476 Potsdam-Golm, Germany. dennie.andresen@ibmt.fraunhofer.de
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: We have successfully shown the OnChip-HDA and applied it for single- and duplex-detection of the pathogens N. gonorrhoeae and S. aureus. CONCLUSION: 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.
BACKGROUND: 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. METHODS: 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. RESULTS: We have successfully shown the OnChip-HDA and applied it for single- and duplex-detection of the pathogens N. gonorrhoeae and S. aureus. CONCLUSION: 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.
Authors: Sebastian Kersting; Valentina Rausch; Frank F Bier; Markus von Nickisch-Rosenegk Journal: Mikrochim Acta Date: 2014-02-18 Impact factor: 5.833
Authors: Claudia Kolm; Roland Martzy; Kurt Brunner; Robert L Mach; Rudolf Krska; Georg Heinze; Regina Sommer; Georg H Reischer; Andreas H Farnleitner Journal: Environ Sci Technol Date: 2017-06-09 Impact factor: 9.028