title: Accurate identification of neutralizing antibodies to adenovirus Ad36, -a putative contributor of obesity in humans.
abstract: BACKGROUND
  In children and adults, human adenovirus serotype 36 (Ad36) is linked with increased adiposity, and important metabolic alterations. Since this property is not shared by many other human adenovirus serotypes, it is imperative to specifically identify exposure to Ad36. Although serum neutralization assay (SNA) is the gold standard to specifically detect neutralizing antibodies (NA) to Ad36, it requires 2-weeks to complete and considerable training to interpret the results. Whereas, an enzyme-immuno assay (EIA) may provide a quicker and objective determination.
  
  
  OBJECTIVE
  Evaluate the accuracy of commercially available EIA kits to detect NA to Ad36. Modify SNA to reduce time and increase objectivity.
  
  
  METHODS
  Sera of 15 seropositive or 16 seronegative subjects confirmed by SNA were used to test: 1) reproducibility of SNA to detect Ad36 exposure, by repeating assays twice; 2) an EIA that detects antibodies to all human adenovirus serotypes (NS-EIA) (Abcam-108705); 3) an EIA supposedly specific for Ad36 antibody (Ad36-EIA) (MyBioSource,#MBS705802), and 4) the concordance of SNA with a novel combination of SNA and immune-staining (SN-IS) kit (Cell BioLabs,#VPK-111).
  
  
  RESULTS
  The SNA showed exact reproducibility. NS-EIA detected adenovirus antibodies in 94% samples, confirming the non-specificity of the assay for Ad36 serotype. All seronegative samples (as determined by SNA) were false positive by Ad36-EIA. In 97% samples, SN-IS showed fidelity with Ad36-antibody status as determined by SNA.
  
  
  CONCLUSIONS
  The available EIA kits are not specific for detecting NA to Ad36. The modified SNA with immune-staining reduces assay time and increases accuracy of detecting by reducing subjectivity.
authors.affiliation: Infections and Obesity Laboratory, Pennington Biomedical Research Center, Baton Rouge, LA 70808 USA.
authors.forename: Olga
authors.initials: O
authors.lastname: Dubuisson
authors.affiliation: Michael & Susan Dell Center for Healthy Living The University of Texas School of Public Health, Houston, TX 77030 USA.
authors.forename: Rena Sue
authors.initials: RS
authors.lastname: Day
authors.affiliation: Infections and Obesity Laboratory, Pennington Biomedical Research Center, Baton Rouge, LA 70808 USA. Electronic address: Nikhil.Dhurandhar@PBRC.EDU.
authors.forename: Nikhil V
authors.initials: NV
authors.lastname: Dhurandhar
mesh_terms: D000256:Adenoviridae / Q000276:immunology* / Q000302:isolation & purification; D000257:Adenoviridae Infections / Q000150:complications* / Q000175:diagnosis; D000328:Adult; D057134:Antibodies, Neutralizing / Q000032:analysis*; D002648:Child; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D009765:Obesity / Q000209:etiology* / Q000503:physiopathology; D012307:Risk Factors; D012680:Sensitivity and Specificity
chemical_list: D057134:Antibodies, Neutralizing
keywords: ELISA; Glycemic control; Infection; Infectobesity; Pediatric obesity