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Apart from its high sensitivity, as compared to other
electrochemical miRNA biosensors, the much-simplified label-
free approach makes the proposed biosensor particularly
attractive for routine miRNA analysis.
30,31
To date, all
miRNA assays must work with highly purified RNA samples.
The freedom from chemical/biological ligation and PCR
amplification greatly improves the suitability in direct profiling
miRNAs with minimal or no sample pretreatment. Attempts
were thus made in utilizing the biosensor for analyzing
circulating miRNAs in blood and miRNAs in total RNA extract
from culture cells. The same sample (except serum) was
simultaneously analyzed by the biosensor and by qPCR. As
presented in Table 1, good agreements between the results
obtained by the biosensor and qPCR on the same sample
confirmed the practical value of the biosensor. In addition, the
results obtained were also consistent with published data.
32,33
Because of its high sensitivity, the amount of RNA needed for asuccessful miRNA detection was in the range of 3−10 ng. In
addition, a direct analysis of let-7 miRNA in serum was
attempted. Unfortunately, persistently lower values with much
larger variations (30%−50%) were obtained. The results were
even worse when RNA extraction efficiency is taken into
consideration. Evidently, the high protein content and other
blood constituents in serum severely interfere with the
detection of miRNA, probably by fouling the biosensor through
nonspecific adsorption. Efforts in developing an antifouling
interface while maintaining most of the attractive features of the
biosensor are underway.
At the present time, this work only serves as a proof-of-
concept through rather labor-intensive manual operation.
Nonetheless, as with other electronic/electrochemical bio-
sensors, both low- and high-density sensor arrays can, in
principle, be constructed by adopting the standard micro-
fabrication technology and the complementary metal-oxide
semiconductor technology that require no additional processes
other than a simple switching mechanism. Two prominent
examples demonstrating the power and capacity of the
electronic/electrochemical detection systems are the electro-
chemical microarray developed by combimatrix
34
and the Ion
Proton Sequencer recently developed by Life Technologies.
35
For example, up to 12 million individal sensors can be
fabricated on a single chip in the Ion Proton Sequencer.
Compared to other label-free electrochemical biosensors,
12−15
the much simplified procedure implies that the throughput can
also be significantly upscaled by upgrading the manual
operation to a microfluidic cartridge. Of course, similar to all
other hybridization-based systems, great care must be taken
when profiling multiple miRNAs sinutaneously, since there is
very little room to fine-tune the hybridization conditions for all
miRNAs.

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