ORIGINAL ARTICLE

Genetic characterisation of adenovirus type 8 isolated in Hiroshima city over a 15 year period

A K Adhikary¹, J Numaga², T Kaburaki², H Kawashima², M Araie², Y Ikeda³, T Ogino³, E Suzuki⁴, H Ushijima⁴, A Mukoyama¹, S Matsuno¹, T Inada¹ and N Okabe¹

¹ Infectious Diseases Surveillance Centre, National Institute of Infectious Diseases, Tokyo 162-8640, Japan
² Department of Ophthalmology, Graduate School of Medicine, The University of Tokyo, Tokyo 114-0024, Japan
³ Hiroshima City Institute of Public Health, Hiroshima 733-8650, Japan
⁴ Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan

Correspondence to:
Correspondence to:
Dr Numaga, Department of Ophthalmology, Graduate School of Medicine, The University of Tokyo, 3-2-1-214, Nishigahara Kita-ku, Tokyo 114-0024, Japan;
jnumaga{at}mub.biglobe.ne.jp

Aims: To investigate the genetic differences among the strains of adenovirus type 8 (Ad8) circulating in Hiroshima city, Japan, and to study their circulation pattern.

Methods: One hundred and twenty nine strains of adenovirus type 8 (Ad8) were isolated in Hiroshima City over a 15 year period (1983–97) from patients with keratoconjunctivitis, and analysed with six restriction enzymes—BamHI, HindIII, PstI, SacI, SalI, and SmaI—to investigate possible relations among the isolates and their genetic variability. Seven hypervariable regions of the hexon gene that carry the type specific epitope were also sequenced to investigate the variation among the genome types.

Results: Restriction endonuclease analyses yielded three known genome types (Ad8A, 13 samples; Ad8B, seven samples; and Ad8E, 35 samples) and a novel genome type (Ad8I, 74 samples). Ad8A, Ad8B, and Ad8E were closely related, with 96% homology, whereas Ad8I had only 71% homology. Ad8A, Ad8B, and Ad8E shared 91.8% and 96.4% homology with regard to their amino acid and nucleotide sequences, respectively, with the isolate 1127 (accession no X74663). However, when compared with Ad8A, Ad8B, Ad8E, and isolate 1127, Ad8I shared only 62.7% and 69.9% homology with regard to amino acid and nucleotide sequences, respectively. Ad8A, Ad8B, and Ad8E had a unique 31 amino acid deletion in the hypervariable region 1 of the hexon gene, whereas Ad8I had a 33 residue deletion. The Ad8E strain that circulated from 1984 to 1995 was stable among the study population. Ad8I was isolated from an outbreak of epidemic keratoconjunctivitis in 1995 and was also isolated from sporadic cases until 1997.

Conclusions: These results confirmed that genetic variability occurs in Ad8 in the microenvironment and revealed the emergence of a new genome type (Ad8I).

Keywords: adenovirus; type 8; genetic characterisation; Hiroshima

Abbreviations: Ad8, adenovirus type 8; EKC, epidemic keratoconjunctivitis; HVR, hypervariable region; PCR, polymerase chain reaction; PCRF, pairwise comigrating restriction fragments; SDS, sodium dodecyl sulfate; TE buffer, Tris/EDTA buffer

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