title: Rapid Production of Virus Protein Microarray Using Protein Microarray Fabrication through Gene Synthesis (PAGES).
abstract: The high genetic variability of RNA viruses is a significant factor limiting the discovery of effective biomarkers, the development of vaccines, and characterizations of the immune response during infection. Protein microarrays have been shown to be a powerful method in biomarker discovery and the identification of novel protein-protein interaction networks, suggesting that this technique could also be very useful in studies of infectious RNA viruses. However, to date, the amount of genetic material required to produce protein arrays, as well as the time- and labor-intensive procedures typically needed, have limited their more widespread application. Here, we introduce a method, protein microarray fabrication through gene synthesis (PAGES), for the rapid and efficient construction of protein microarrays particularly for RNA viruses. Using dengue virus as an example, we first identify consensus sequences from 3,604 different strains and then fabricate complete proteomic microarrays that are unique for each consensus sequence. To demonstrate their applicability, we show that these microarrays can differentiate sera from patients infected by dengue virus, related pathogens, or from uninfected patients. We anticipate that the microarray and expression library constructed in this study will find immediate use in further studies of dengue virus and that, more generally, PAGES will become a widely applied method in the clinical characterization of RNA viruses.
authors.affiliation: From the ‡Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine, Ministry of Education.
authors.forename: Huan
authors.initials: H
authors.lastname: Qi
authors.affiliation: §Institute of Pathogenic Microbiology, Center for Disease Control and Prevention of Guangdong, No. 160, Quanxian Road, Dashi Street, Panyu District, Guangzhou, 511430, China.
authors.forename: Huiqiong
authors.initials: H
authors.lastname: Zhou
authors.affiliation: ¶School of Biomedical Engineering.
authors.forename: Daniel Mark
authors.initials: DM
authors.lastname: Czajkowsky
authors.affiliation: From the ‡Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine, Ministry of Education.
authors.forename: Shujuan
authors.initials: S
authors.lastname: Guo
authors.affiliation: From the ‡Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine, Ministry of Education.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.affiliation: From the ‡Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine, Ministry of Education.|‖National Research Institute for Health and Family Planning, No.12 Dahuisi Road, Haidian District, Beijing, 100081, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: From the ‡Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine, Ministry of Education.
authors.forename: Yi
authors.initials: Y
authors.lastname: Shi
authors.affiliation: §Institute of Pathogenic Microbiology, Center for Disease Control and Prevention of Guangdong, No. 160, Quanxian Road, Dashi Street, Panyu District, Guangzhou, 511430, China.
authors.forename: Lifeng
authors.initials: L
authors.lastname: Lin
authors.affiliation: From the ‡Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine, Ministry of Education.
authors.forename: Jingfang
authors.initials: J
authors.lastname: Wang
authors.affiliation: §Institute of Pathogenic Microbiology, Center for Disease Control and Prevention of Guangdong, No. 160, Quanxian Road, Dashi Street, Panyu District, Guangzhou, 511430, China.|**State Key Laboratory of Oncogenes and Related Genes,Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, China.
authors.forename: De
authors.lastname: Wu
authors.affiliation: From the ‡Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine, Ministry of Education, taosc@sjtu.edu.cn wu_de68@cdcp.org.cn.|¶School of Biomedical Engineering.|**State Key Laboratory of Oncogenes and Related Genes,Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, China.
authors.forename: Sheng-Ce
authors.initials: SC
authors.lastname: Tao
mesh_terms: D000328:Adult; D000368:Aged; D003715:Dengue / Q000097:blood / Q000821:virology*; D003716:Dengue Virus / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015723:Gene Library; D014644:Genetic Variation; D006801:Humans; D008297:Male; D008875:Middle Aged; D010802:Phylogeny; D040081:Protein Array Analysis / Q000379:methods*; D060066:Protein Interaction Maps; D040901:Proteomics / Q000379:methods; D014764:Viral Proteins / Q000032:analysis* / Q000097:blood* / Q000235:genetics; D055815:Young Adult
chemical_list: D014764:Viral Proteins