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Backfilling rolling cycle amplification with enzyme-DNA conjugates on antibody for portable electrochemical immunoassay with glucometer readout
Lilin Ge1,2; Bin Li1,2; Houxi Xu1; Wenyuan Pu1; Hang Fai Kwok2
2019-05
Source PublicationBIOSENSORS & BIOELECTRONICS
ISSN0956-5663
Volume132Pages:210-216
AbstractA simple and feasible electrochemical immunosensing protocol with glucometer readout was designed for the detection of low-abundance disease-related biomarker (alpha-fetoprotein; AFP) on the basis of backfilling rolling cycle amplification (RCA) with invertase-DNA2 conjugates on the detection antibody. The assay consisted of the immunoreaction, RCA reaction, DNA2-invertase hybridization and glucose measurement. Initially, a sandwiched immunoreaction was carried out between anti-AFP capture antibody-coated microplate between nanogold-labeled pAb2 detection antibody conjugated with DNA1 primer (DNA1-AuNP-pAb2) in the presence of target ATP. Thereafter, the carried primers triggered the RCA reaction in the presence of circular DNA template, polymerase and dNTP, to produce numerous repeated oligonucleotide sequences for hybridization with many invertase-DNA2 conjugates. The carried invertase molecules accompanying the hybridization reaction hydrolyzed sucrose into glucose, thereby resulting in the amplification of the detectable signal on a handheld personal glucometer (PGM). Under optimum conditions, the developed immunoassay exhibited high sensitivity for the quantitative screening of AFP within a dynamic range of 0.1–100 ng mL−1 at a low detection limit of 0.087 ng mL−1. Other biomarkers and proteins did not interfere the signals of this system. In addition, this method was utilized to determine human serum samples containing target AFP, and received well-matched results with the referenced enzyme-linked immunosorbent assay (ELISA) method.
KeywordElectrochemical Immunoassayv Rolling Cycle Amplification Glucometer Readout Enzyme-dna Conjugates Disease-related Biomarker
DOI10.1016/j.bios.2019.02.051
Indexed BySCI
Language英语
WOS Research AreaBiophysics ; Biotechnology & Applied Microbiology ; Chemistry ; Electrochemistry ; Science & Technology - Other Topics
WOS SubjectBiophysics ; Biotechnology & Applied Microbiology ; Chemistry, Analytical ; Electrochemistry ; Nanoscience & Nanotechnology
WOS IDWOS:000464296300024
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Cited Times [WOS]:6   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionFaculty of Health Sciences
Affiliation1.State Key Laboratory Cultivation Base for TCM Quality and Efficacy, Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Qixia District, Nanjing 210046, PR China
2.Institute of Translational Medicine, Faculty of Health Sciences, University of Macau, Avenida de Universidade, Taipa, PR China
First Author AffilicationFaculty of Health Sciences
Recommended Citation
GB/T 7714
Lilin Ge,Bin Li,Houxi Xu,et al. Backfilling rolling cycle amplification with enzyme-DNA conjugates on antibody for portable electrochemical immunoassay with glucometer readout[J]. BIOSENSORS & BIOELECTRONICS,2019,132:210-216.
APA Lilin Ge,Bin Li,Houxi Xu,Wenyuan Pu,&Hang Fai Kwok.(2019).Backfilling rolling cycle amplification with enzyme-DNA conjugates on antibody for portable electrochemical immunoassay with glucometer readout.BIOSENSORS & BIOELECTRONICS,132,210-216.
MLA Lilin Ge,et al."Backfilling rolling cycle amplification with enzyme-DNA conjugates on antibody for portable electrochemical immunoassay with glucometer readout".BIOSENSORS & BIOELECTRONICS 132(2019):210-216.
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