title: Comparison of E and NS1 antigens capture ELISA to detect dengue viral antigens from mosquitoes.
abstract: OBJECTIVE
  In the absence of an effective vaccine or specific antiviral therapy against dengue infection, the only available control measure remains focusing on the incrimination and reduction of vector (mosquito) populations to suppress virus transmission. Diagnosis of dengue in laboratory can be carried out using several approaches, however, their sensitivity and specificity vary from test-to-test. This study was conducted to evaluate the sensitivity and stability of viral envelope (E) and NS1 antigens detected by ELISA in dengue virus infected mosquitoes.
  
  
  METHODS
  An in-house developed E-ELISA to detect dengue E antigens was first characterized by using cross-reactive monoclonal antibody (mAb) 42-3 and rabbit polyclonal antibodies as the capture and detector antibodies, respectively. The sensitivity of E-ELISA was compared with the Platelia Dengue NS1 Ag kit using experimentally infected or field-caught mosquitoes.
  
  
  RESULTS
  Our results demonstrated that the E-ELISA was capable of detecting viral antigens with the sensitivity of 69.57, 100, 52.38 and 66.67% for DENV-1 to DENV-4 infected mosquito pools, respectively. This was comparable to the Platelia Dengue NS1 Ag kit, detecting 100% of DENV-1 infected mosquito pools. Among 124 field-collected mosquito pools collected in the vicinity of localized outbreak areas; both E-ELISA and NS1 Ag kit confirmed nine RT-PCR positive samples with sensitivity and concordance rate up to 100%.
  
  
  CONCLUSIONS
  With the future potential of antigen capture ELISA to be used in the resource deprived regions, the study showed that E-ELISA has similar sensitivity and antigen stability as NS1 Ag kit to complement the current established virological surveillance in human. The improvement of the sensitivity in detecting DENV-3/4 will be needed to incorporate this method into routine mosquito surveillance system.
authors.affiliation: Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung-Hsing University, Taichung, Taiwan.
authors.forename: Day-Yu
authors.initials: DY
authors.lastname: Chao
authors.forename: Yi-Jung
authors.initials: YJ
authors.lastname: Liu
authors.forename: Wen-Fan
authors.initials: WF
authors.lastname: Shen
authors.forename: Wu-Chun
authors.initials: WC
authors.lastname: Tu
authors.forename: Jedhan Ucat
authors.initials: JU
authors.lastname: Galula
authors.forename: Han-Chung
authors.initials: HC
authors.lastname: Wu
mesh_terms: D000330:Aedes / Q000821:virology*; D000818:Animals; D000914:Antibodies, Viral / Q000302:isolation & purification; D000956:Antigens, Viral / Q000302:isolation & purification*; D002460:Cell Line; D003716:Dengue Virus / Q000145:classification / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D065288:Serogroup; D014759:Viral Envelope Proteins / Q000302:isolation & purification*; D017361:Viral Nonstructural Proteins / Q000302:isolation & purification*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; C072281:E protein TH Sman, Dengue virus