TY - JOUR
T1 - Robust label-free microRNA detection using one million ISFET array
AU - Ganguli, Anurup
AU - Watanabe, Yoshihiko
AU - Hwang, Michael T.
AU - Huang, Jui Cheng
AU - Bashir, Rashid
PY - 2018/6/2
Y1 - 2018/6/2
N2 - Detection of nucleic acid molecules is one of the most pervasive assays in biology, medicine, and agriculture applications. Currently, most comely used DNA/RNA detection platforms use fluorescence labeling and require lab-scale setting for performing the assay. There is a need for developing less expensive, label-free, and rapid detection of biomolecules with minimal utilization of resources. Use of electrical approaches for detection of biomolecules by utilizing their inherent charge is a promising direction for biosensing assays. Here, we report a 1024 × 1024 array of Ion Sensitive Field Effect Transistors (ISFET) as label free sensors for detection of nucleic acid molecules. Using PNA probe functionalized on these ISFET array, we robustly detected miRNA Let-7b by measuring changes in drain current after hybridization of target molecules with concentration as low as 1 nM. We demonstrate that mismatched or non-complementary target molecules resulted in statistically smaller changes. Most importantly, the high-density sensor array shows unprecedented reliability and robustness with P values <0.0001 for all experiments. Practical implementation of this platform could have a wide range of applications in high-throughput nucleic acid genotyping, detection of amplified pathogenic nucleic acid, detection of cell-free DNA, and electrical readouts for current hybridization-based DNA biomolecular assays.
AB - Detection of nucleic acid molecules is one of the most pervasive assays in biology, medicine, and agriculture applications. Currently, most comely used DNA/RNA detection platforms use fluorescence labeling and require lab-scale setting for performing the assay. There is a need for developing less expensive, label-free, and rapid detection of biomolecules with minimal utilization of resources. Use of electrical approaches for detection of biomolecules by utilizing their inherent charge is a promising direction for biosensing assays. Here, we report a 1024 × 1024 array of Ion Sensitive Field Effect Transistors (ISFET) as label free sensors for detection of nucleic acid molecules. Using PNA probe functionalized on these ISFET array, we robustly detected miRNA Let-7b by measuring changes in drain current after hybridization of target molecules with concentration as low as 1 nM. We demonstrate that mismatched or non-complementary target molecules resulted in statistically smaller changes. Most importantly, the high-density sensor array shows unprecedented reliability and robustness with P values <0.0001 for all experiments. Practical implementation of this platform could have a wide range of applications in high-throughput nucleic acid genotyping, detection of amplified pathogenic nucleic acid, detection of cell-free DNA, and electrical readouts for current hybridization-based DNA biomolecular assays.
KW - 1024 × 1024 array
KW - Electrical DNA sensor
KW - Electrical microarray
KW - ISFET
KW - Multiplexed biosensing
UR - http://www.scopus.com/inward/record.url?scp=85056666044&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85056666044&partnerID=8YFLogxK
U2 - 10.1007/s10544-018-0290-8
DO - 10.1007/s10544-018-0290-8
M3 - Article
C2 - 29858718
AN - SCOPUS:85056666044
VL - 20
JO - Biomedical Microdevices
JF - Biomedical Microdevices
SN - 1387-2176
IS - 2
ER -