TY - JOUR
T1 - Accurate identification of neutralizing antibodies to adenovirus Ad36, -a putative contributor of obesity in humans
AU - Dubuisson, Olga
AU - Day, Rena Sue
AU - Dhurandhar, Nikhil V.
N1 - Funding Information:
Funding: Funding for the study was provided in part by the ‘Assistance to Firefighters Grants program’ managed by the Federal Emergency Management Agency in the Department of Homeland Security (EMW- Dhurandhar, 2011, 2012 -FP-01971) and (EMW-2010-FP-01812). This study was presented as a poster during the ‘Obesity Week 2013’ meeting held in Atlanta, GA.
PY - 2015/1/1
Y1 - 2015/1/1
N2 - 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. Objectives Evaluate the accuracy of commercially available EIA kits to detect NA to Ad36. Modify SNA to reduce time and increase objectivity. Study Design 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.
AB - 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. Objectives Evaluate the accuracy of commercially available EIA kits to detect NA to Ad36. Modify SNA to reduce time and increase objectivity. Study Design 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.
KW - ELISA
KW - Glycemic control
KW - Infection
KW - Infectobesity
KW - Pediatric obesity
UR - http://www.scopus.com/inward/record.url?scp=84920512595&partnerID=8YFLogxK
U2 - 10.1016/j.jdiacomp.2014.09.004
DO - 10.1016/j.jdiacomp.2014.09.004
M3 - Article
C2 - 25312598
AN - SCOPUS:84920512595
SN - 1056-8727
VL - 29
SP - 83
EP - 87
JO - Journal of Diabetes and its Complications
JF - Journal of Diabetes and its Complications
IS - 1
ER -