title: An ultrasensitive electrogenerated chemiluminescence-based immunoassay for specific detection of Zika virus.
abstract: Zika virus (ZIKV) is a globally emerging mosquito-transmitted flavivirus that can cause severe fetal abnormalities, including microcephaly. As such, highly sensitive, specific, and cost-effective diagnostic methods are urgently needed. Here, we report a novel electrogenerated chemiluminescence (ECL)-based immunoassay for ultrasensitive and specific detection of ZIKV in human biological fluids. We loaded polystyrene beads (PSB) with a large number of ECL labels and conjugated them with anti-ZIKV monoclonal antibodies to generate anti-ZIKV-PSBs. These anti-ZIKV-PSBs efficiently captured ZIKV in solution forming ZIKV-anti-ZIKV-PSB complexes, which were subjected to measurement of ECL intensity after further magnetic beads separation. Our results show that the anti-ZIKV-PSBs can capture as little as 1 PFU of ZIKV in 100 μl of saline, human plasma, or human urine. This platform has the potential for development as a cost-effective, rapid and ultrasensitive assay for the detection of ZIKV and possibly other viruses in clinical diagnosis, epidemiologic and vector surveillance, and laboratory research.
authors.affiliation: Department of Biological Sciences, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
authors.forename: Dhiraj
authors.initials: D
authors.lastname: Acharya
authors.affiliation: Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
authors.forename: Pradip
authors.initials: P
authors.lastname: Bastola
authors.affiliation: Department of Biological Sciences, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
authors.forename: Linda
authors.initials: L
authors.lastname: Le
authors.affiliation: Department of Biological Sciences, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
authors.forename: Amber M
authors.initials: AM
authors.lastname: Paul
authors.affiliation: Departments of Medicine, Molecular Microbiology, Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Estefania
authors.initials: E
authors.lastname: Fernandez
authors.affiliation: Departments of Medicine, Molecular Microbiology, Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Diamond
authors.affiliation: Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
authors.forename: Wujian
authors.initials: W
authors.lastname: Miao
authors.affiliation: Department of Biological Sciences, University of Southern Mississippi, Hattiesburg, MS 39406, USA.
authors.forename: Fengwei
authors.initials: F
authors.lastname: Bai
mesh_terms: D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry*; D000914:Antibodies, Viral / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D008163:Luminescent Measurements / Q000379:methods*; D012680:Sensitivity and Specificity; D000071244:Zika Virus; D000071243:Zika Virus Infection / Q000097:blood* / Q000175:diagnosis
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000914:Antibodies, Viral