Helicase dependent OnChip-amplification and its use in multiplex pathogen detection.
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.


CONCLUSIONS
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.
Andresen
Dennie
D
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
von Nickisch-Rosenegk
Markus
M
Bier
Frank F
FF
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D004265:DNA Helicases / Q000378:metabolism*; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification*; D046228:Microarray Analysis; D046208:Microchip Analytical Procedures / Q000379:methods*; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D046915:Penicillin-Binding Proteins
D001426:Bacterial Proteins; D046915:Penicillin-Binding Proteins; C487789:mecA protein, Staphylococcus aureus; D004265:DNA Helicases