title: Ultra-sensitive chemiluminescence imaging DNA hybridization method in the detection of mosquito-borne viruses and parasites.
abstract: Mosquito-borne viruses (MBVs) and parasites (MBPs) are transmitted through hematophagous arthropods-mosquitoes to homoiothermous vertebrates. This study aims at developing a detection method to monitor the spread of mosquito-borne diseases to new areas and diagnose the infections caused by MBVs and MBPs.
  
  
  
  In this assay, an ultra-sensitive chemiluminescence (CL) detection method was developed and used to simultaneously detect 19 common MBVs and MBPs. In vitro transcript RNA, virus-like particles (VLPs), and plasmids were established as positive or limit of detection (LOD) reference materials.
  
  
  
  MBVs and MBPs could be genotyped with high sensitivity and specificity. The cut-off values of probes were calculated. The absolute LODs of this strategy to detect serially diluted in vitro transcribed RNAs of MBVs and serially diluted plasmids of MBPs were 102-103copies/μl and 101-102copies/μl, respectively. Further, the LOD of detecting a strain of pre-quantified JEV was 101.8-100.8PFU/ml, fitted well in a linear regression model (coefficient of determination = 0.9678).
  
  
  
  Ultra-sensitive CL imaging DNA hybridization was developed and could simultaneously detect various MBVs and MBPs. The method described here has the potential to provide considerable labor savings due to its ability to screen for 19 mosquito-borne pathogens simultaneously.
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing, 100850, People's Republic of China.|Postdoctoral Research Workstation, 210th Hospital of the Chinese People's Liberation Army, Dalian, 116021, People's Republic of China.
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing, 100850, People's Republic of China.|Beijing Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Beijing, 100850, People's Republic of China.
authors.forename: Qiqi
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing, 100850, People's Republic of China.|Beijing Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Beijing, 100850, People's Republic of China.
authors.forename: Biao
authors.initials: B
authors.lastname: Zhou
authors.affiliation: Postdoctoral Research Workstation, 210th Hospital of the Chinese People's Liberation Army, Dalian, 116021, People's Republic of China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Wang
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing, 100850, People's Republic of China. doudoucsh@sina.com.|Beijing Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Beijing, 100850, People's Republic of China. doudoucsh@sina.com.
authors.forename: Suhong
authors.initials: S
authors.lastname: Chen
authors.affiliation: Beijing Institute of Radiation Medicine, Beijing, 100850, People's Republic of China. sqwang@bmi.ac.cn.|Beijing Key Laboratory of New Molecular Diagnosis Technologies for Infectious Diseases, Beijing, 100850, People's Republic of China. sqwang@bmi.ac.cn.
authors.forename: Shengqi
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D009033:Culicidae / Q000469:parasitology* / Q000821:virology*; D008163:Luminescent Measurements / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010271:Parasites / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
keywords: Chemiluminescence; DNA hybridization; Mosquito-borne parasites; Mosquito-borne viruses