TY - JOUR
T1 - An ultrasensitive electrogenerated chemiluminescence-based immunoassay for specific detection of Zika virus
AU - Acharya, Dhiraj
AU - Bastola, Pradip
AU - Le, Linda
AU - Paul, Amber M.
AU - Fernandez, Estefania
AU - Diamond, Michael S.
AU - Miao, Wujian
AU - Bai, Fengwei
N1 - Funding Information:
We thank Dr. Barbara W. Johnson (CDC Arbovirus Branch, Fort Collins CO) for providing ZIKV, Dr. Robert B. Tesh (University of Texas Medical Branch) for providing CHIKV, and Dr. John F. Anderson at the Connecticut Agricultural Experiment Station for providing WNV and dengue viruses. We thank Dr. Jennifer Givero and Derek Platt for technical help with generating antibodies. We also thank Mississippi-IDeA Network of Biomedical Research Excellence (MS-INBRE) for the use of research facility. This work was supported in part by financial support from the National Science Foundation CAREER award (CHE-0955878, W.M.) and National Institute of Allergy and Infectious Diseases of the National Institutes of Health (R15AI113706 to F.B. and R01 AI073755 toM.S.D.).
PY - 2016/8/24
Y1 - 2016/8/24
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84984691524&partnerID=8YFLogxK
U2 - 10.1038/srep32227
DO - 10.1038/srep32227
M3 - Article
C2 - 27554037
AN - SCOPUS:84984691524
SN - 2045-2322
VL - 6
JO - Scientific reports
JF - Scientific reports
M1 - 32227
ER -