Development of a standard protocol for quantitative polymerase chain reaction to detect adenovirus 36, which is associated with obesity

K. A. Hwang, S. Park, J. H. Ahn, J. H. Nam

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

It has been previously reported that adenovirus 36 (Ad36) infection is associated with obesity in humans and other animals. However, there is no clinically available standard protocol to detect Ad36 DNA. In this study, we developed a method for quantitative and rapid detection of Ad36 DNA. Using a TaqMan probe quantitative polymerase chain reaction (qPCR), we identified that the E3 and E4orf1 regions specifically detect Ad36 DNA, because these regions did not show cross reactivity with other types of adenoviruses. The limit of detection was 379 copy/ml and 384 copy/ml for E3 and E4orf1 regions of Ad36, respectively. The %CV (coefficient of variation) for reproducibility of the assay using adenovirus reference material ranged from 1.07-13.02. After we developed the standard protocol to detect Ad36 DNA, we used mouse as a surrogate model to confirm its clinical applicability. We administered Ad36 to mice via intranasal and oral routes, with intraperitoneal administration as the positive control, to analyze the effect of infection route. Ad36 DNA could be detected in lungs, liver, pancreas, and epididymal fat tissue after intraperitoneal injection, whereas it was found only in lungs after intranasal injection. No Ad36 DNA was detectable in any tested organ after oral injection. This indicates that the main route of infection with Ad36 is intranasal, suggesting that Ad36 is a respiratory virus. The standard protocol for qPCR developed in this study is useful for clinical detection of Ad36 DNA.

Original languageEnglish
Pages (from-to)350-359
Number of pages10
JournalActa Virologica
Volume62
Issue number4
DOIs
Publication statusPublished - 2018
Externally publishedYes

Bibliographical note

Funding Information:
Acknowledgments. This work was supported by the Catholic University of Korea, Research Fund, 2018, a grant from the Korean Health Technology R&D Project through the KHIDI, funded by the Ministry of Health & Welfare, Republic of Korea (HI15C2955), a grant (16172MFDS363) from Ministry of Food and Drug Safety in 2017–2018, and Basic Science Research Program through the NRF, which is funded by the Ministry of Science, ICT & Future Planning (NRF-2015M3A9B5030157). All authors are the guarantors for this work, and had full access to all of the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis. We thank the researchers at Catholic University Viral Immunology Lab and Genetree Research Company for helpful discussions and their technical assistance.

Publisher Copyright:
© 2018 AEPress s.r.o. All rights reserved.

Keywords

  • Adenovirus 36
  • Obesity
  • Real-time PCR

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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