Archives of Virology

, Volume 151, Issue 8, pp 1587–1602 | Cite as

Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene

  • X. Lu
  • D. D. Erdman


Human adenoviruses (Ads) are responsible for a substantial disease burden. Type-specific identification of Ads can help guide therapeutic and disease prevention strategies and aid epidemiological investigations. Immunotyping of Ads by serum neutralization (SN) is laborious and time consuming and depends upon type-specific antisera that are in short supply. A rapid molecular typing assay based on polymerase chain reaction (PCR) amplification and sequencing of Ad hexon gene hyper-variable regions 1–6 (HVR1–6) known to contain type-specific epitopes was evaluated as an alternative to SN. Deduced amino acid sequences of HVR1–6 obtained from all 51 currently recognized Ad prototype strains were well resolved, with the exception of types 15 and 29, which were identical. Of 192 temporally and geographically diverse Ad field isolates sequenced in this study, and 111 previously published sequences, all more closely matched their predicted prototype strains. Ads were also detected and correctly identified directly from 24 clinical specimens positive by culture or antigen detection. PCR and sequencing of HVR1–6 offers a practical alternative to SN for typing most Ads and can be readily adapted for use in laboratories with molecular capabilities.


Molecular Typing Acute Respiratory Disease Human Adenovirus Prototype Strain Hexon 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. Adhikary, AK, Numaga, J, Kaburaki, T, Kawashima, H, Kato, S, Araie, M, Miyata, K, Shimizu, H, Yagyu, F, Suzuki, E, Ushijima, H 2001Rapid detection and typing of oculopathogenic Strain of subgenus D adenoviruses by fiber-based PCR and restriction enzyme analysisInvest Opthi Vis Sci4220102015Google Scholar
  2. Adhikary, AK, Inada, T, Numaga, J, Suzuki, E, Ushijima, H, Banik, U, Mukouyama, A, Matsuno, S, Okabe, N 2004Characterization of hexon and fiber genes of a novel strain of adenovirus involved in epidemic kerato-conjunctivitisJ Clin Pathol579597PubMedCrossRefGoogle Scholar
  3. Adrian, T, Wadell, G, Hierholzer, JC, Wigand, R 1986DNA restriction analysis of adenovirus prototypes 1 to 41Arch Virol91277290PubMedCrossRefGoogle Scholar
  4. Ariga, T, Shimada, Y, Ohgami, K 2004New genome type of adenovirus serotype 4 caused nosocomial infections associated with epidemic conjunctivitis in JapanJ Clin Microbiol4236443648PubMedCrossRefGoogle Scholar
  5. Athappily, FK, Murali, R, Rux, JJ, Cai, Z, Burnett, RM 1994The refined crystal structure of hexon, the major coat protein of adenovirus type 2, at 2.9 A resolutionJ Mol Biol242430455CrossRefGoogle Scholar
  6. Benkö, M, Harrach, B, Russell, WC 2000Adenoviridae. Family Adenoviridaevan Regenmortel, MHVFauquet, CMBishop, DHLCarstens, EBEstes, MKLemon, SMManiloff, JMayo, MAMcGeoch, DJPringle, CRWickner, RB eds. Virus taxonomyAcademic PressSan Diego, CA227238Google Scholar
  7. Benson, DA, Karsch-Mizrachi, I, Lipman, DJ, Ostell, J, Rapp, BA, Wheeler, DL 2002GenBankNucleic Acids Res301720PubMedCrossRefGoogle Scholar
  8. Blasiole, DA, Metzgar, D, Daum, LT, Ryan, MA, Wu, J, Wills, C, Le, CT, Freed, NE, Hansen, CJ, Gray, GC, Russell, KL 2004Molecular analysis of adenovirus isolates from vaccinated and unvaccinated young adultsJ Clin Microbiol4216861693PubMedCrossRefGoogle Scholar
  9. Boursnell, MEG, Mautner, V 1981Recombination in adenovirus: crossover sites in intertypic recombinants are located in regions of homologyVirology112198209PubMedCrossRefGoogle Scholar
  10. Crawford-Miksza, L, Schnurr, DP 1996Analysis of 15 adenovirus hexon proteins reveals the location and structure of seven hypervariable regions containing serotype-specific residuesJ Virol7018361844PubMedGoogle Scholar
  11. Crawford-Miksza, L, Schnurr, DP 1996Adenovirus serotype evolution is driven by illegitimate recombination in the hypervariable regions of the hexon proteinVirology224357367PubMedCrossRefGoogle Scholar
  12. Crawford-Miksza, L, Nang, RN, Schnurr, DP 1999Strain variation in adenovirus serotypes 4 and 7a causing acute respiratory diseaseJ Clin Microbiol3711071112PubMedGoogle Scholar
  13. De Jong, JC, Wermenbol, AG, Verweij-Uijterwaal, MW, Slaterus, KW, Wertheim-Van Dillen, P, Van Doornum, GJ, Khoo, SH, Hierholzer, JC 1996Adenovirus from AIDS patients, including two new candidate serotypes Ad50 and Ad51 of subgeneus D and B1, respectivelyXth International Congress of VirologyJerusalem, Israel230Google Scholar
  14. Gaydos, CA, Gaydos, JC 1995Adenovirus vaccines in the U.S. militaryMil Med160300304PubMedGoogle Scholar
  15. Gruber, WC, Russell, DJ, Tibbetts, C 1993Fiber gene and genomic origin of human adenovirus type 4Virology196603611PubMedCrossRefGoogle Scholar
  16. Hierholzer, JC, Bingham, PG 1978Vero microcultures for adenovirus neutralization testsJ Clin Microbiol7499506PubMedGoogle Scholar
  17. Hierholzer, JC 1995AdenovirusesLennette, EHLennette, DALennette, ET eds. Diagnostic procedures for viral, rickettsial, and chlamydial infections7American Public Health AssociationWashington DC169188Google Scholar
  18. Hierholzer, JC, Suggs, MT 1969Standardized viral hemagglutination and hemagglutination-inhibition tests. I. Standardization of erythrocyte suspensionsAppl Microbiol18816823PubMedGoogle Scholar
  19. Hierholzer, JC, Suggs, MT, Hall, EC 1969Standardized viral hemagglutination and hemagglutination-inhibition tests. II. Description and statistical evaluationAppl Microbiol18824833PubMedGoogle Scholar
  20. Hierholzer, JC, Gamble, WC, Dowdle, WR 1975Reference equine antisera to 33 human adenovirus types: homologous and heterologous titersJ Clin Microbiol16574PubMedGoogle Scholar
  21. Hierholzer, JC, Pumarola, A 1976Antigenic characterization of intermediate adenovirus 14-11 strains associated with upper respiratory illness in a military campInfect Immunol13354359Google Scholar
  22. Hierholzer, JC, Wigand, R, Anderson, LJ, Adrian, T, Gold, JW 1988Adenoviruses from patients with AIDS: a plethora of serotypes and a description of five new s serotypes of subgenus D (types 43–47)J Infect Dis58804813Google Scholar
  23. Hierholzer, JC, Stone, YO, Broderson, JR 1991Antigenic relationships among the 47 human adenoviruses determined in reference horse antiseraArch Virol121179197PubMedCrossRefGoogle Scholar
  24. Kidd, AH, Jonsson, M, Garwicz, D, Kajon, AE, Wermenbol, AG, Verweij, MW, De Jong, JC 1996Rapid subgenus identification of human adenovirus isolates by a general PCRJ Clin Microbiol34622627PubMedGoogle Scholar
  25. Li, L, Shimizu, H, Doan, LT, Tung, PG, Okitsu, S, Nishio, O, Suzuki, E, Seo, JK, Kim, KS, Muller, WE, Ushijima, H 2004Characterization of adenovirus type 41 isolates from children with acute gastroenteritis in Japan, Vietnam, and KoreaJ Clin Microbiol4240324039PubMedCrossRefGoogle Scholar
  26. Lin, B, Vora, GJ, Thach, D 2004Use of oligonucleotide microarrays for rapid detection and serotyping of acute respiratory disease-associated adenovirusesJ Clin Microbiol4232323239PubMedCrossRefGoogle Scholar
  27. Lukashok, SA, Horwitz, MS 1998New perspectives in adenovirusesCurr Clin Top Infect Dis18286305PubMedGoogle Scholar
  28. Morfin, F, Dupuis-Girod, S, Mundweiler, S, Falcon, D, Carrington, D, Sedlacek, P, Bierings, M, Cetkovsky, P, Kroes, AC, van Tol, MJ, Thouvenot, D 2005In vitro susceptibility of adenovirus to antiviral drugs is species-dependentAntivir Ther10225229PubMedGoogle Scholar
  29. Pring-Åkerblom, P, Adrian, T, Kostler, T 1997PCR-based detection and typing of human adenoviruses in clinical samplesRes Virol148225231PubMedCrossRefGoogle Scholar
  30. Pring-Åkerblom, P, Adrian, T 1993The hexon genes of adenoviruses of subgenus C: comparison of the variable regionsRes Virol144117127PubMedGoogle Scholar
  31. Pring-Åkerblom, P, Adrian, T 1994Type- and group-specific polymerase chain reaction for adenovirus detectionRes Virol1452535PubMedGoogle Scholar
  32. Pring-Åkerblom, P, Trijssenaar, FE, Adrian, T 1995Sequence characterization and comparison of human adenovirus subgenus B and E hexonsVirology212232236PubMedCrossRefGoogle Scholar
  33. Rocholl, C, Gerber, K, Daly, J, Pavia, AT, Byington, CL 2004Adenoviral infections in children: the impact of rapid diagnosisPediatrics1135156CrossRefGoogle Scholar
  34. Saitoh-Inagawa, W, Oshima, A, Aoki, K 1996Rapid diagnosis of adenovirus conjunctivitis by PCR and restriction fragment length polymorphism analysisJ Clin Microbiol3421132116PubMedGoogle Scholar
  35. Sarantis, H, Johnson, G, Brown, M, Petric, M, Tellier, R 2004Comprehensive detection and serotypeing of human adenoviruses by PCR and sequencingJ Clin Microbiol4230633069CrossRefGoogle Scholar
  36. Schnurr, D, Dondero, ME 1993Two new candidate adenovirus serotypesIntervirology367983PubMedGoogle Scholar
  37. Shimada, Y, Ariga, T, Yoshitsugu, T, Aoki, K, Ohno, S, Ishiko, H 2004Molecular diagnosis of human adenoviruses D and E by a phylogeny-based classification method using a partial hexon sequenceJ Clin Microbiol4215771584PubMedCrossRefGoogle Scholar
  38. Singh-Naz, N, Rodriguez, WJ, Kidd, AH, Brandt, CD 1988Monoclonal antibody enzyme-linked immunosorbent assay for specific identification and typing of subgroup F adenovirusesJ Clin Microbiol26297300PubMedGoogle Scholar
  39. Simmons, MP, Ochoterena, H 2000Gaps as characters in sequence-based phylogenetic analysesSyst Biol49369381PubMedCrossRefGoogle Scholar
  40. Swofford DL (2003) PAUP*. Phylogenetic Analysis Using Parsimony (*and other methods), 4th edn. Sinauer Associates, Sunderland, MassachusettsGoogle Scholar
  41. Takeuchi, S, Itoh, N, Uchio, E, Aoki, K, Ohno, S 1999Serotyping of adenoviruses on conjunctival scrapings by PCR and sequence analysisJ Clin Microbiol3718391845PubMedGoogle Scholar
  42. Takeuchi, S, Itoh, N, Uchio, E, Tanaka, K, Kitamura, N, Kanai, H, Isobe, K, Aoki, K, Ohno, S 1999Adenovirus strains of subgenus D associated with nosocomial infection as new etiological agents of epidemic keratoconjunctivitis in JapanJ Clin Microbiol3733923394PubMedGoogle Scholar
  43. Wigand, R 1987Pitfalls in the identification of adenovirusesJ Virol Methods16161169PubMedCrossRefGoogle Scholar
  44. Wigand, R, Sehn, N, Hierholzer, JC, de Jong, JC, Adrian, T 1985Immunological and biochemical characterization of human adenoviruses from subgenus B. I. Antigenic relationshipsArch Virol846378PubMedCrossRefGoogle Scholar
  45. Wood, SR, Sharp, IR, Caul, EO 1997Rapid detection and serotyping of adenovirus by direct immuno-fluorescenceJ Med Virol51198201PubMedCrossRefGoogle Scholar
  46. Xu, W, McDonough, MM, Erdman, DD 2000Species-specific identification of human adenoviruses by a multiplex polymerase chain reaction assayJ Clin Microbiol3841144120PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • X. Lu
    • 1
  • D. D. Erdman
    • 1
  1. 1.Division of Viral and Rickettsial DiseasesCenters for Disease Control and PreventionAtlantaU.S.A.

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