caa a22 4500 606155937 CHVBK 20210128100606.0 cr unu---uuuuu 210128e20150701xx s 000 0 eng 10.1007/s10096-015-2353-6 doi (NATIONALLICENCE)springer-10.1007/s10096-015-2353-6 A multiplex ELISA-based protein array for screening diagnostic antigens and diagnosis of Flaviviridae infection [Elektronische Daten] [D. Wang, Y. Zheng, X. Kang, X. Zhang, H. Hao, W. Chen, L. Liu, X. Li, L. Li, Q. Yuan, F. Chen, Y. Yang, Y. Jiang, H. Jiang] Assays with the ability to detect multiple antibodies in parallel have a wide range of potential applications in epidemiologic research. Here, a multiplex enzyme-linked immunosorbent assay-based protein array (ELISA-array) was developed to simultaneously detect five Flaviviridae infections. The platform was based on an indirect ELISA and 15 antigens were constructed for specific antibody detection against five Flaviviridae viruses (Japanese B, tick-borne encephalitis, West Nile, dengue, and yellow fever viruses) and four serotypes of dengue virus. The specificity was evaluated by calculating the signal value cross-reacting with serum immunized with other viruses, and the sensitivity of antigens was compared with conventional ELISAs using immunized rabbit polyclonal antisera. IgG and IgM calibration curves were constructed to evaluate the reproducibility of the platform. Finally, 24 dengue fever (DF) infection and 15 tick-borne encephalitis (TBE) infection clinical sera were used to compare the advantage of ELISA-array to ELISA. After initial screening, 9 out of 15 antigens were chosen for ELISA-array printing. By using different virus-immunized rabbit antiserum, 7 out of 9 antigens showed good specificity in the ELISA-array. Eight out of 9 antigens showed four-fold greater sensitivity in ELISA-array compared to that in conventional ELISAs. The coefficients of determination (r 2) close to 1 showed high reproducibility, and clinical sera test showed that ELISA-array had higher specificity and sensitivity than traditional ELISA. ELISA-array was a good platform for antigen screening and this multiplexed assay might be a useful and convenient tool for multiple immunological detection of infectious viral antibodies. Springer-Verlag Berlin Heidelberg, 2015 Wang D. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Zheng Y. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Kang X. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Zhang X. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Hao H. CAS Key Laboratory of Pathogenic Microbiology and Immunity, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut Chen W. Genomics Institute in Shenzhen, Beishan Industry Beishan Road, 518083, Shenzhen, China aut Liu L. Genomics Institute in Shenzhen, Beishan Industry Beishan Road, 518083, Shenzhen, China aut Li X. Genomics Institute in Shenzhen, Beishan Industry Beishan Road, 518083, Shenzhen, China aut Li L. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Yuan Q. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Chen F. College of Food Technology, Huazhong Agricultural University, Wuhan, China aut Yang Y. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Jiang Y. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut Jiang H. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/7(2015-07-01), 1327-1336 0934-9723 34:7<1327 2015 34 10096 https://doi.org/10.1007/s10096-015-2353-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10096-015-2353-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang D. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Zheng Y. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Kang X. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Zhang X. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Hao H. CAS Key Laboratory of Pathogenic Microbiology and Immunity, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China aut NATIONALLICENCE 700 1- Chen W. Genomics Institute in Shenzhen, Beishan Industry Beishan Road, 518083, Shenzhen, China aut NATIONALLICENCE 700 1- Liu L. Genomics Institute in Shenzhen, Beishan Industry Beishan Road, 518083, Shenzhen, China aut NATIONALLICENCE 700 1- Li X. Genomics Institute in Shenzhen, Beishan Industry Beishan Road, 518083, Shenzhen, China aut NATIONALLICENCE 700 1- Li L. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Yuan Q. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Chen F. College of Food Technology, Huazhong Agricultural University, Wuhan, China aut NATIONALLICENCE 700 1- Yang Y. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Jiang Y. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 700 1- Jiang H. State Key Laboratory of Pathogen and Biosecurity, Academy of Military Medical Sciences, 100071, Beijing, China aut NATIONALLICENCE 773 0- European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/7(2015-07-01), 1327-1336 0934-9723 34:7<1327 2015 34 10096