Microfluidic analysis of multiplex,,,PCR products for the genotyping of,,,Helicobacter pylori
nd 2 of figure 2A aredisplayed as electropherograms(figures 2B and 2C). Finally combined microfluidicanalysis of all 7 alleles of H.pylori amplified in 4 differentmPCRs was possible on the chipafter mixing all mPCR reactionsprior to analysis (combination of:iceA1/A2 vacA s1/s2 cagA vacAm1/m2). This is difficult wi
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.
