First Identification of Human Adenovirus Subtype 21a in China With MinION and Illumina Sequencers

Ye, Fuqiang; Han, Yifang; Zhu, Juanjuan; Li, Peng; Zhang, Qi; Lin, Yanfeng; Wang, Taiwu; Lv, Heng; Wang, Changjun; Wang, Chunhui; Zhang, Jinhai

A throat swab sample was collected from a 20-year-old male outpatient with fever and sore throat in March 2019. The swab sample was immediately placed into preservation solution (Yocon, Beijing, China) and stored at -80 • C until further processing. Initial real-time PCR tests of viral respiratory pathogens including HAdV, coronavirus, and influenza A/B viruses obtained a positive result for HAdV by using universal primers. However, PCR primers for epidemic HAdV subtypes in China, including HAdV1, HAdV3, HAdV4, HAdV7, HAdV14, and HAdV55, displayed negative results. The sample was cultivated in HEp-2 cell lines (ATCC, Manassas, VA, United States) supplemented with Dulbecco modified eagle medium (Life Technologies, Carlsbad, CA, United States) containing 2% fetal bovine serum (Life Technologies) until 90% cytopathic effects were visible. The patient provided written informed consent upon enrollment. The study conformed to the ethical guidelines of the 1975 Declaration of Helsinki and was approved by the Institutional Review Board of the Center for Disease Control and Prevention of Eastern Theater Command.