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Microfluidic analysis of multiplex,,,PCR products for the genotyping of,,,Helicobacter pylori

nd 2 of figure 2A are displayed as electropherograms (figures 2B and 2C).

Finally, combined microfluidic analysis of all 7 alleles of H. pylori amplified in 4 different mPCRs was possible on the chip after mixing all mPCR reactions prior to analysis (combination of: iceA1/A2, vacA s1/s2, cagA, vacA m1/m2). This is difficult with slab gel electrophoresis where differences in the amount of PCR products cause difficulties in data interpretation. All seven different mPCR products within a range of 102-301 bp could be separated and quantified (figure 3). These results reveal that Lab-on-a-chip technology enables the separation of seven different targets restricted to a size range up to 300 bp in a combined mPCR reaction.

Conclusion
The results of this study clearly demonstrate the ability of the Agilent 2100 bioanalyzer to differentiate the allelic H. pylori variations, vacAs1/s2/cagA, of vacAm1/m2 and of iceA1/2, amplified by mPCR. The high resolution achieved by the microfluidic based Lab-on-the-Chip technology allows to analyze a broad and expanding panel of virulence and risk factors. This is of great importance since an extended spectrum of prognostic or therapeutically relevant information used in H. pylori diagnostics will now be accessible for routinely processed simultaneous diagnostics by means of mRCR assays.



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