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2014 ; 6
(20
): 8173-8179
Nephropedia Template TP
gab.com Text
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English Wikipedia
MICROFLUIDIC DEVICES FOR LABEL-FREE AND NON-INSTRUMENTED QUANTITATION OF
UNAMPLIFIED NUCLEIC ACIDS BY FLOW DISTANCE MEASUREMENT
#MMPMID25530814
Chatterjee D
; Mansfield DS
; Woolley AT
Anal Methods
2014[Oct]; 6
(20
): 8173-8179
PMID25530814
show ga
Timely biomarker quantitation has potential to improve human health but current
methods have disadvantages either in terms of cost and complexity for benchtop
instruments, or reduced performance in quantitation and/or multiplexing for
point-of-care systems. We previously developed microfluidic devices wherein
visually observed flow distances correlated with a model analyte's
concentration.(1) Here, we significantly expand over this prior result to
demonstrate the measurement of unamplified DNA analogues of microRNAs (miRNAs),
biomarkers whose levels can be altered in disease states. We have developed a
method for covalently attaching nucleic acid receptors on poly(dimethylsiloxane)
microchannel surfaces by silane and cross-linker treatments. We found a flow
distance dependence on target concentrations from 10 ?g/mL to 10 pg/mL for DNA in
both buffer and synthetic urine. Moreover, flow time in addition to flow distance
is correlated with target concentration. We also observed longer flow distances
for single-base mismatches compared to the target sequence at the same
concentration, indicating that our approach can be used to detect point
mutations. Finally, experiments with DNA analogues of miRNA biomarkers for kidney
disease (mir-200c-3p) and prostate cancer (mir-107) in synthetic urine showed the
ability to detect these analytes near clinically relevant levels. Our results
demonstrate that these novel microfluidic assays offer a simple route to
sensitive, amplification-free nucleic acid quantitation, with strong potential
for point-of-care application.
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