TY - JOUR
T1 - Adenovirus 36 antibody detection
T2 - Improving the standard serum neutralization assay
AU - Chappell, Cynthia L.
AU - Dickerson, Mary
AU - Day, R. Sue
AU - Dubuisson, Olga
AU - Dhurandhar, Nikhil V.
N1 - Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - Adenovirus 36 (AdV36) causes weight gain in animal models, including non-human primates. In humans, AdV36-neutralizing antibodies are associated with adiposity; however, longitudinal studies in large populations are needed to clarify AdV36's contribution. The current gold standard for detection of AdV36-specific antibody is the serum neutralization assay (SNA), which requires long incubation times and highly trained personnel. The standard SNA was modified using an immunocytochemical (ICC) approach, which allows for a more rapid and objective assessment of AdV36 antibodies. Using the ICC assay, virus-infected cells were detected as early as day 1 (D1) and by D5 were detected in 100% of microtiter wells versus 20.3% of wells detected by observing the cytopathic effect. Further, human sera tested with the ICC assay at D5 had a sensitivity and specificity of 80.0% and 95.7%, respectively, when compared to the standard SNA read at D11. Thus, the ICC assay decreased assay incubation time, provided a more objective and easily interpreted assessment, and had a high degree of sensitivity and specificity in determining serological status. The more rapid and objective ICC method will make large population studies feasible, improve comparability among laboratories, and contribute to understanding the role of AdV36 in obesity.
AB - Adenovirus 36 (AdV36) causes weight gain in animal models, including non-human primates. In humans, AdV36-neutralizing antibodies are associated with adiposity; however, longitudinal studies in large populations are needed to clarify AdV36's contribution. The current gold standard for detection of AdV36-specific antibody is the serum neutralization assay (SNA), which requires long incubation times and highly trained personnel. The standard SNA was modified using an immunocytochemical (ICC) approach, which allows for a more rapid and objective assessment of AdV36 antibodies. Using the ICC assay, virus-infected cells were detected as early as day 1 (D1) and by D5 were detected in 100% of microtiter wells versus 20.3% of wells detected by observing the cytopathic effect. Further, human sera tested with the ICC assay at D5 had a sensitivity and specificity of 80.0% and 95.7%, respectively, when compared to the standard SNA read at D11. Thus, the ICC assay decreased assay incubation time, provided a more objective and easily interpreted assessment, and had a high degree of sensitivity and specificity in determining serological status. The more rapid and objective ICC method will make large population studies feasible, improve comparability among laboratories, and contribute to understanding the role of AdV36 in obesity.
KW - Adiposity
KW - Elisa
KW - Immunochemical staining
KW - Infectobesity
KW - Obesity
KW - Serological assay
UR - http://www.scopus.com/inward/record.url?scp=85006886586&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2016.10.014
DO - 10.1016/j.jviromet.2016.10.014
M3 - Article
C2 - 27825952
AN - SCOPUS:85006886586
SN - 0166-0934
VL - 239
SP - 69
EP - 74
JO - Journal of Virological Methods
JF - Journal of Virological Methods
ER -