title: Simple and cost-effective restriction endonuclease analysis of human adenoviruses.
abstract: Restriction endonuclease analyses (REAs) constitute the only inexpensive molecular approach capable of typing and characterizing human adenovirus (HAdV) strains based on the entire genome. However, the application of this method is limited by the need for time-consuming and labor-intensive procedures. We herein developed a simple and cost-effective REA for assessing HAdV. The method consists of (1) simple and cost-effective DNA extraction, (2) fast restriction endonuclease (RE) digestion, and (3) speedy mini agarose gel electrophoresis. In this study, DNA was isolated according to the kit-based method and 21.0 to 28.0  μg of viral DNA was extracted from prototypes (HAdV-1, HAdV-3, HAdV-4, and HAdV-37) in each flask. The amount of DNA ranged from 11.4 to 57.0  μg among the HAdV-3 (n=73) isolates. The obtained viral DNA was found to be applicable to more than 10 types of REAs. Fast-cut restriction endonucleases (REs) were able to digest the DNA within 15 minutes, and restriction fragments were easily separated via horizontal mini agarose gel electrophoresis. The whole procedure for 10 samples can be completed within approximately six hours (the conventional method requires at least two days). These results show that our REA is potentially applicable in many laboratories in which HAdVs are isolated.
authors.affiliation: Unit of Microbiology, Faculty of Medicine, AIMST University, Semeling, 08100 Bedong, Kedah Darul Aman, Malaysia ; Infectious Disease Surveillance Center, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
authors.forename: Arun Kumar
authors.identifier: 0000-0002-6009-1608
authors.initials: AK
authors.lastname: Adhikary
authors.affiliation: Infectious Disease Surveillance Center, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
authors.forename: Nozomu
authors.initials: N
authors.lastname: Hanaoka
authors.affiliation: Infectious Disease Surveillance Center, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
authors.forename: Tsuguto
authors.identifier: 0000-0002-4861-4349
authors.initials: T
authors.lastname: Fujimoto
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics*; D045744:Cell Line, Tumor; D004262:DNA Restriction Enzymes / Q000737:chemistry* / Q000191:economics; D004279:DNA, Viral / Q000737:chemistry; D004587:Electrophoresis, Agar Gel / Q000191:economics / Q000379:methods; D016678:Genome; D016679:Genome, Viral; D006801:Humans; D000091402:Prohibitins; D015183:Restriction Mapping / Q000379:methods*; D013045:Species Specificity
chemical_list: D004279:DNA, Viral; C000717331:PHB2 protein, human; D000091402:Prohibitins; D004262:DNA Restriction Enzymes
references.citation: Clin Microbiol Rev. 1999 Oct;12(4):612-26
references.pmid: 10515905
references.citation: J Clin Microbiol. 1988 May;26(5):1026-8
references.pmid: 2838502
references.citation: Methods Mol Biol. 1991;7:109-28
references.pmid: 21416352
references.citation: Emerg Infect Dis. 2002 Mar;8(3):269-77
references.pmid: 11927024
references.citation: Jpn J Infect Dis. 2011;64(6):493-8
references.pmid: 22116328
references.citation: Biotechniques. 1997 May;22(5):868-70
references.pmid: 9149866
references.citation: J Med Microbiol. 2012 Nov;61(Pt 11):1491-1503
references.pmid: 22956755
references.citation: Jpn J Infect Dis. 2014;67(4):282-7
references.pmid: 25056074
references.citation: Ann N Y Acad Sci. 1980;354:16-42
references.pmid: 6261642
references.citation: J Clin Pathol. 2003 Feb;56(2):120-5
references.pmid: 12560390
references.citation: Adv Exp Med Biol. 2010;659:153-74
references.pmid: 20204763
references.citation: Leukemia. 2010 Apr;24(4):706-14
references.pmid: 20147979
references.citation: Microbiol Immunol. 1983;27(9):817-22
references.pmid: 6316118
references.citation: J Med Virol. 1996 Feb;48(2):151-6
references.pmid: 8835348
references.citation: J Med Virol. 2009 Aug;81(8):1380-4
references.pmid: 19551823
references.citation: J Clin Microbiol. 2004 Dec;42(12):5489-92
references.pmid: 15583271
references.citation: J Clin Microbiol. 1988 May;26(5):1009-15
references.pmid: 2838500
references.citation: J Gen Virol. 1985 Dec;66 ( Pt 12):2685-91
references.pmid: 2999311
references.citation: J Clin Microbiol. 2003 Oct;41(10):4594-9
references.pmid: 14532188
references.citation: Arch Virol. 1986;91(3-4):277-90
references.pmid: 3022681
references.citation: Acta Paediatr. 1993 Jun-Jul;82(6-7):557-61
references.pmid: 8338989
references.citation: J Mol Biol. 1967 Jun 14;26(2):365-9
references.pmid: 4291934
references.citation: J Med Microbiol. 2001 Jun;50(6):571-574
references.pmid: 11393295
references.citation: J Clin Microbiol. 2001 Feb;39(2):498-505
references.pmid: 11158096
references.citation: Jpn J Infect Dis. 2010 Sep;63(5):338-41
references.pmid: 20859000
references.citation: J Clin Microbiol. 2005 Feb;43(2):726-32
references.pmid: 15695671
references.citation: J Clin Virol. 2009 Dec;46(4):331-6
references.pmid: 19854101
references.citation: J Virol. 2011 Jun;85(11):5701-2
references.pmid: 21450823