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. 2017 Dec 14;12(12):e0189625.
doi: 10.1371/journal.pone.0189625. eCollection 2017.

Whole-genome analyses of human adenovirus type 55 emerged in Tibet, Sichuan and Yunnan in China, in 2016

Wenbo Wang  1 , Yuan Liu  2 , Yifan Zhou  1 , Liangqi Gu  1 , Lin Zhang  1 , Xuelian Zhang  1 , Maomao Chen  1 , Ziying Zou  2 , Wei Qiu  1 , Xiaobing Hu  1 , Quanshui Fan  1
Affiliations

Whole-genome analyses of human adenovirus type 55 emerged in Tibet, Sichuan and Yunnan in China, in 2016

Wenbo Wang et al. PLoS One. .

Abstract

Three outbreaks of acute respiratory disease occurred at military camps in 2016 at Tibet, Sichuan and Yunnan province, China. The pathogen induced these three outbreaks were all confirmed as HAdV-55 by genotype-specific PCR. The outbreak in Tibet was the first report that HAdV-55 occurred in the high altitude (HA, above sea level 3658 m). This study aims to determine the gene variation and evolution characteristics of these viral strains. Three strains of adenoviruses, LS89/Tibet/2016 (GenBank accession no. KY002683), SF04/SC/2016 (GenBank accession no. KY002684) and KM03/YN/2016 (GenBank accession no. KY002685) were obtained and confirmed by wholegenome sequencing. No multi-gene fragments recombination were found in these isolated HAdV-55 virus compared with previous reported HAdV-55 strains in China. The outbreaks in Tibet and in Sichuan continuously occurred. Virus isolated from Tibet (LS89/Tibet/2016) and Sichuan (SF04/SC/2016) had a similar mutation pattern and had a closer genetic evolutionary distance than KM03/YN/2016 strain, which indicates that the pathogens causing these two outbreaks may be of the same origin. Moreover, we found that heating was an effective way to inactive these viruses, which provide valuable information for the development of HAdV-55 vaccines. Our data provide new information for genetic evolution of HAdV-55, and contribute to the prevention and control of HAdV-55 infection in the future.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Phylogenetic analysis of isolated HAdV strains.
Phylogenetic tree of human adenovirus based on the whole genome sequence was constructed by neighbor-joining method with 1000 bootstrap replicates. The strains isolated in our study are labeled as bold text.
Fig 2
Fig 2. Variation sites based on the China/QS-DLL/2006 strain.
The black line represents the mutation, the red line represents base insertion, and the green line represents base deletion.
Fig 3
Fig 3. Phylogenetic analysis of HAdV-55 strains.
Phylogenetic tree of HAdV-55 strains based on the whole genome sequence was constructed by neighbor-joining method with 1000 bootstrap replicates. The strains isolated in our study are labeled as bold text.
Fig 4
Fig 4. Viral growth kinetics of HAdV-B55 in HEp-2 cells.
HEp-2 cells were infected with 100 CCID50 HAdV-B55 viruses. The supernatants and cells were separately harvested at 1, 2, 3, 5 and 6 d post-infection. The extracellular viral loads in the supernatants (A) and the intracellular viral loads in the cells (B) were determined by qPCR. The data are shown as the mean 6 standard deviation of three independent experiments. (C) Toxicity of HAdV-B55 in HEp-2 cells. Data were shown the means and standard errors of three replicate assays. WT, XZ, SC and YN present HAdV-55 strains of Y16/SX/2011, LS89/Tibet/2016, SF04/SC/2016 and KM03/YN/2016 respectively.
Fig 5
Fig 5. The effect of heat and ultraviolet treatment on the infectivity of HAdV-55.
The viruses were incubated for 30 minutes at 56°C or treated for ultraviolet irradiation at a wavelength of 254 nm for 30 minutes. (A) Viral DNA levels after treatment were determined by qPCR. (B) Viral titers in heat- or ultraviolet- treated virus samples were determined by infection on Hep-2 cells. Data were shown the means and standard errors of three replicate assays (* P < 0.05, compared with control). WT, XZ, SC and YN present HAdV-55 strains of Y16/SX/2011, LS89/Tibet/2016, SF04/SC/2016 and KM03/YN/2016 respectively.
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Grants and funding

This study was funded by PLA Logistics Research Program (BWS14J025), PLA Medical Science Youth Training Project (14QNP057), National Natural Science Foundation of China (81301445), the 12th Five Years Major Special Projects of Chinese PLA (AWS11L009) and Open Research Fund Program of Shanghai Key Laboratory of Medical Biodefense (SKLM1401). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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