title: Development of a highly sensitive gold nanoparticle probe-based assay for bluetongue virus detection.
abstract: A simple gold nanoparticle (GNP) probe based assay (GNPA) that was modified from a bio-barcode assay (BCA) technique, was developed for ultra-sensitive, rapid detection of the bluetongue virus (BTV) VP7 outer-core protein. This assay captures the VP7 target antigen using the GNP probe coated with anti-VP7 polyclonal antibodies and single-stranded signal DNA. Magnetic microparticle (MMP) probes coated with anti-VP7 monoclonal antibodies were then added to form a sandwich immuno-complex. The single-stranded signal DNA coated onto the GNP probe present in the immuno-complex could then be detected by PCR and real-time fluorescence PCR using a TaqMan probe. The assay has a purified VP7 detection limit of 10(-2)fg/ml which is 8 orders of magnitude greater than that of conventional antigen capture ELISAs and 1 order of magnitude more sensitive than that of a conventional BCA. These results indicate that the GNPA is a highly sensitive method for easy detection of BTV proteins and that it can be modified as needed to measure the presence of other proteins.
authors.affiliation: Viral Safety Laboratory of the National Center of Biomedical Analysis, Institute of Transfusion Medicine, The Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Hui-qiong
authors.initials: HQ
authors.lastname: Yin
authors.forename: Min-xian
authors.initials: MX
authors.lastname: Jia
authors.forename: Shu
authors.initials: S
authors.lastname: Yang
authors.forename: Pei-pei
authors.initials: PP
authors.lastname: Jing
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.forename: Jin-gang
authors.initials: JG
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000914:Antibodies, Viral; D000956:Antigens, Viral / Q000032:analysis; D001819:Bluetongue / Q000175:diagnosis* / Q000821:virology; D001820:Bluetongue virus / Q000302:isolation & purification*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D007118:Immunoassay / Q000379:methods; D053758:Nanoparticles; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D012756:Sheep; D014730:Veterinary Medicine / Q000379:methods*; D014758:Viral Core Proteins / Q000032:analysis*; D014773:Virology / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D014758:Viral Core Proteins; C066862:VP7 protein, orbivirus