TY - JOUR
T1 - Sensitive detection of motor neuron disease derived exosomal miRNA using electrocatalytic activity of gold-loaded superparamagnetic ferric oxide nanocubes
AU - Masud, Mostafa Kamal
AU - Mahmudunnabi, Rabbee G.
AU - Aziz, Nahian Binte
AU - Stevens, Claire H.
AU - Do-Ha, Dzung
AU - Yang, Shu
AU - Blair, Ian P.
AU - Hossain, Md Shahriar A.
AU - Shim, Yoon Bo
AU - Ooi, Lezanne
AU - Yamauchi, Yusuke
AU - Shiddiky, Muhammad J. A.
PY - 2020/8/17
Y1 - 2020/8/17
N2 - Dysregulated microRNA associated pathways contribute to the pathology of neurological disorders, hence presenting themselves as a potential candidate for motor neuron disease (MND) diagnosis. Herein, we reported an enzymatic amplification-free approach for the electrochemical detection of exosomal microRNA (miR-338-3p) from preconditioned media of motor neurons obtained from amyotrophic lateral sclerosis (ALS) patients and healthy controls. Our assay utilizes a three-step strategy that involves i) initial isolation and purification of exosomal miR-338-3p from patients and healthy controls using biotinylated complementary capture probe followed by heat-release of the specific target, ii) direct adsorption of target miR-338-3p onto the gold-loaded ferric oxide nanocatalyst (AuNP-Fe2O3NC) through affinity interaction between microRNA and exposed gold surfaces within the AuNP-Fe2O3NC, and iii) gold nanocatalyst-induced electrocatalytic signal amplification through methylene blue-ferricyanide redox cycling (MB/[Fe(CN)6]3−). The electrocatalytic signal is monitored by using chronocoulometry at the AuNP–Fe2O3NC-modified screen-printed carbon electrode (AuNP-Fe2O3NC/SPCE). We demonstrated the detection of miR-338-3p as low as 100 aM in spiked buffer samples with a relative standard deviation of (%RSD) <5.0 % (n=5). We also demonstrate the successful detection of miR-338-3p from a small cohort of preconditioned media of motor neurons obtained from ALS patients and healthy controls. The sensor avoids the use of conventional recognition and transduction layers in hybridization-based electrochemical miRNA biosensors, polymerase-based amplifications. It is robust, fast (<2.5 h) and potentially applicable to a wide variety of RNA biomarker detection.
AB - Dysregulated microRNA associated pathways contribute to the pathology of neurological disorders, hence presenting themselves as a potential candidate for motor neuron disease (MND) diagnosis. Herein, we reported an enzymatic amplification-free approach for the electrochemical detection of exosomal microRNA (miR-338-3p) from preconditioned media of motor neurons obtained from amyotrophic lateral sclerosis (ALS) patients and healthy controls. Our assay utilizes a three-step strategy that involves i) initial isolation and purification of exosomal miR-338-3p from patients and healthy controls using biotinylated complementary capture probe followed by heat-release of the specific target, ii) direct adsorption of target miR-338-3p onto the gold-loaded ferric oxide nanocatalyst (AuNP-Fe2O3NC) through affinity interaction between microRNA and exposed gold surfaces within the AuNP-Fe2O3NC, and iii) gold nanocatalyst-induced electrocatalytic signal amplification through methylene blue-ferricyanide redox cycling (MB/[Fe(CN)6]3−). The electrocatalytic signal is monitored by using chronocoulometry at the AuNP–Fe2O3NC-modified screen-printed carbon electrode (AuNP-Fe2O3NC/SPCE). We demonstrated the detection of miR-338-3p as low as 100 aM in spiked buffer samples with a relative standard deviation of (%RSD) <5.0 % (n=5). We also demonstrate the successful detection of miR-338-3p from a small cohort of preconditioned media of motor neurons obtained from ALS patients and healthy controls. The sensor avoids the use of conventional recognition and transduction layers in hybridization-based electrochemical miRNA biosensors, polymerase-based amplifications. It is robust, fast (<2.5 h) and potentially applicable to a wide variety of RNA biomarker detection.
KW - amyotrophic lateral sclerosis (ALS)
KW - electrocatalytic activity
KW - electrochemical detection
KW - microRNA
KW - motor neuron
KW - nanotechnology
UR - http://www.scopus.com/inward/record.url?scp=85089961386&partnerID=8YFLogxK
UR - http://purl.org/au-research/grants/arc/DP190102944
UR - http://purl.org/au-research/grants/arc/FT150100479
UR - http://purl.org/au-research/grants/nhmrc/1135720
U2 - 10.1002/celc.202000828
DO - 10.1002/celc.202000828
M3 - Article
AN - SCOPUS:85089961386
SN - 2196-0216
VL - 7
SP - 3459
EP - 3467
JO - ChemElectroChem
JF - ChemElectroChem
IS - 16
ER -