title: An improved gold nanoparticle probe-based assay for HCV core antigen ultrasensitive detection.
abstract: A gold nanoparticle probe-based assay (GNPA) was developed for ultrasensitive detection of Hepatitis C virus (HCV) core antigen. In the GNPA, after anti-HCV core antigen polyclonal antibodies and single-stranded barcode signal DNA were labeled on gold nanoparticle probe (NP), DNA enzyme was used to degrade the unbound barcode DNAs. The anti-HCV core antigen monoclonal antibodies were coated on magnetic microparticles probe (MMP). Then the NP-HCV core antigen-MMP sandwich immuno-complex was formed when the target antigen protein was added and captured. Magnetically separated, the immuno-complex containing the single-stranded barcode signal DNA was characterized by TaqMan probe based real-time fluorescence PCR. A detection limit of 1 fg/ml was determined for the HCV core antigen which is magnitude greater than that of ELISA (2ng/ml). The coefficients of variation (CV) of intra-assay and inter-assay respectively ranged from 0.22-2.62% and 1.92-3.01%. The improved GNPA decreased the interference of unbound barcode DNAs and may be an new way for HCV core antigen detection.
authors.affiliation: Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Hui-Qiong
authors.initials: HQ
authors.lastname: Yin
authors.affiliation: Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Chang-Fu
authors.initials: CF
authors.lastname: Ji
authors.affiliation: Beijing Institute of Basic Medical Science, Beijing 100850, China.
authors.forename: Xi-Qin
authors.initials: XQ
authors.lastname: Yang
authors.affiliation: Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.affiliation: Beijing Institute of Transfusion Medicine, Beijing 100850, China.
authors.forename: Shu
authors.initials: S
authors.lastname: Yang
authors.affiliation: Beijing Institute of Basic Medical Science, Beijing 100850, China.
authors.forename: He-Qiu
authors.initials: HQ
authors.lastname: Zhang
authors.affiliation: Beijing Institute of Transfusion Medicine, Beijing 100850, China. Electronic address: zhangjg@nic.bmi.cn.
authors.forename: Jin-Gang
authors.initials: JG
authors.lastname: Zhang
mesh_terms: D000956:Antigens, Viral / Q000032:analysis*; D006046:Gold; D016174:Hepacivirus / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D053758:Nanoparticles; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014758:Viral Core Proteins / Q000032:analysis*
chemical_list: D000956:Antigens, Viral; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus; D006046:Gold
keywords: Bio-bar code assay; Gold nanoparticle probe based assay; Hepatitis C virus; Ultrasensitive detection