Investigation of Adenoviruses in Patients with Conjunctivitis by Various Methods

Aslı Kiraz 1, Selma Gökahmetoğlu 2, Osman Özüberk 2, Ayşe Öner 3
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1 Çanakkale Onsekiz Mart University, Faculty of Medicine, Department of Microbiology and Clinical Microbiology, Çanakkale, Turkey
2 Erciyes University, Faculty of Medicine, Department of Medical Microbiology, Kayseri, Turkey
3 Erciyes University, Faculty of Medicine, Department of Ophthalmology, Kayseri, Turkey
EUR J BASIC MED SCI, Volume 3, Issue 4, pp. 83-89.
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Adenoviruses are the most common causative agents of epidemics of community acquired and nosocomial conjunctivitis. Adenoviruses are resistant to physical and chemical factors and they can spread easily. Due to these factors, rapid and accurate diagnosis is very important in adenoviral eye infections. In this study, it was aimed to investigate adenoviruses from conjunctival swab samples of 50 patients suspected of adenoviral conjunctivitis by two different cell culture and PCR methods. Conjunctival swab samples from patients suspected of adenoviral conjunctivitis were taken between March 2012- December 2012. Adenovirus DNA was investigated by real-time PCR method in specimens. Samples were inoculated on conventional HEp-2 cell culture and HEp-2 shellvial cell culture. Virus isolation in conventional HEp-2 cell culture was confirmed with an immunofluorescence kit and a monoclonal antibody kit. Adenovirus DNA was found positive in 23 (46 %) of 50 conjunctival samples by real-time PCR. Virus isolation in conventional HEp-2 cell culture was shown in 19 (38 %) samples by immunofluorescence method and 13 (26 %) samples by monoclonal antibody method. Results obtained from Argene immunofluorescence kit were considered as positive results for conventional HEp-2 cell culture method. Shell-vial culture could be performed in 24 swab samples and 17 of samples were found positive. These 17 samples that found positive in shell-vial culture were also positive with PCR and conventional HEp-2 cell culture methods. Additionally, four samples were found positive by PCR but adenovirus could not be isolated by two cell culture methods. Two samples that were found positive with PCR and conventional HEp-2 cell culture methods were found negative by shell-vial cell culture. Considering HEp-2 cell culture confirmed with immunofluorescence method as gold standart, sensitivities and specifities were found as 100 % and 87 % respectively for PCR, 89 % and 100 % respectively for shellvial cell culture. The cell culture method is still important in adenoviral eye infections but can not provide rapid diagnosis. In adenoviral eye infections, it is considered that real-time PCR is a more useful method that provides results in the shortest period of time with high sensitivity.


Kiraz A, Gökahmetoğlu S, Özüberk O, Öner A. Investigation of Adenoviruses in Patients with Conjunctivitis by Various Methods. Eur J Basic Med Sci. 2013;3(4):83-9.


  • Ishiko H, Shimada Y, Konno T, et al. Novel human adenovirus causing nosocomial epidemic keratoconjunctivitis. J Clin Microbiol 2008;46(6):2002-8.
  • Damen M, Minnaar R, Glasius P, et al. Real-time PCR with an internal control for detection of all known human adenovirus serotypes. J Clin Microbiol 2008;46(12):3997-4003.
  • Buckwalter SP, Teo R, Espy MJ, Sloan LM, Smith TF, Pritt BS. Real-time qualitative PCR for 57 human adenovirus types from multiple specimen sources. J Clin Microbiol 2012;50(3):766-71.
  • Ishiko H, Aoki K. Spread of epidemic keratoconjunctivitis due to a novel serotype of human adenovirus in Japan. J Clin Microbiol 2009;47(8):2678-9.
  • Maranhao AG, Soares CC, Albuquerque MC, Santos N. Molecular epidemiology of adenovirus conjunctivitis in Rıo de Janeıro, Brazil, between 2004 and 2007. Rev Inst Med Trop S Paulo 2009;51(4):227-9.
  • Butt AL, Chodosh J. Adenoviral keratoconjunctivitis in a tertiary care eye clinic. Cornea 2006;25(2):199-202.
  • Nakamura M, Hirano E, Kowada K, et al. Surveillance of adenovirus D in patients with epidemic keratoconjunctivitis from Fukui prefecture,Japan,1995-2010. J Med Virol 2012;84:81-6.
  • Matsui K, Shimizu H, Yoshida A, Nagaoka E, Nishio O, Okuda K. Monitoring of adenovirus from conjunctival scrapings in Japan during 2005-2006. J Med Virol 2008;80:997-1003.
  • Meyer-Rüsenberg B, Loderstadt U, Richard G, Kaulfers P, Gesser C. Epidemic keratoconjunctivitis. Dtsch Arztebl Int 2011;108(27):475-80.
  • Echavarria M. Adenoviruses in immuncompromised hosts. Clin Microbiol Rev 2008;21(4):704-15.
  • Echavarria M. Adenoviruses. In: Zuckerman AJ, Banatvala JE, Schoub BD, Griffiths PD, Mortimer P (eds), Principles&Practice of Clinical Virology (6th ed) Wiley-Blackwell Press, UK 2009, 463-89.
  • Percivalle E, Sarasini A, Torsellini M, et al. A comparison of methods for detecting adenovirus type 8 keratoconjunctivitis during a nosocomial outbreak in a neonatal intensive care unit. J Clin Virol 2003;28:257-64.
  • Levent F, Greer JM, Snider M, Demmler-Harrison GJ. Performance of a new immunochromatographic assay for detection of adenoviruses in children. J Clin Virol 2009;44:173-5.
  • Kaneko H, Mori S, Suzuki O, et al. The cotton rat model for adenovirus ocular infection:antiviral activity of cidofovir. Antiviral Research 2004;61:63-6.
  • Mahmood AR, Narang AT. Diagnosis and management of the acute red eye. Emerg Med Clin N Am 2008;26:35-55.
  • Lernout T, Maillard O, Boireaux S, Collet L, Filleul L. A large outbreak of conjunctivitis on Mayotte Island, France, February to May 2012. Eurosurveill 2012;17:23.
  • Hovding G. Acute bacterial conjunctivitis. Acta Ophthalmol 2008;86:5-17.
  • Kinchington PR, Romanowski EG, Gordon YJ. Prospects for adenovirus antivirals. J Antimicrob Chemother 2005;55:424-9.
  • Yağcı R, Akçalı A, Yağcı S, et al. Molecular identification of adenoviral conjunctivitis in Turkey. Eur J Ophthalmol 2010;20(4):669-74.
  • Robinson C, Echavarria M. Adenoviruses. In: Versalovic J, Carroll KC, Funke G, Jorgensen JH, Landry ML, Warnock DW (eds), Manual of Clinical Microbiology (10th ed). ASM Press, Washington 2011.
  • Schrauder A, Altmann D, Laude G, et al. Epidemic conjunctivitis in Germany,2004. Eurosurveill 2006;11:185-7.
  • Wölfel R, Pfeffer M, Essbauer S, Nerkelun S, Dobler G. Evaluation of sampling technique and transport media for the diagnostics of adenoviral eye infections. Graefe’s Arch Clin Exp Ophthalmol 2006;244:1497-504.
  • Lu X, Erdman DD. Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene. Arch Virol 2006;151:1587-602.
  • Cheung D, Bremner J, Chan JTK. Epidemic keratoconjunctivitis-do outbreaks have to be epidemic? Eye 2003;17:356-63.
  • Jin X, Ishii A, Aoki K, Ishida S, Mukasa K, Ohno S. Detection of human adenovirus hexon antigen using carbon nanotube sensors. J Virol Meth 2011;171:405-7.
  • Tabbara KF, Omar N, Hammouda E, et al. Molecular epidemiology of adenoviral keratoconjunctivitis in Saudi Arabia. Mol Vis 2010;16:2132-6.
  • Huang M, Nguy L, Ferrenberg J, Boeckh M, Cent A, Corey L. Development of multiplexed real-time quantitative PCR assay for detecting human adenoviruses. Diagn Microbiol Infect Dis 2008;62(3):263-71.
  • Brooks GF, Carroll KC, Butel JS, et al (eds). Jawetz, Melnick&Adelberg’s Medical Microbiology (26th ed). McGraw-Hill Press, USA 2013.
  • Kojaoghlanian T, Flomenberg P, Horwitz MS. The impact of adenovirus infection on the immuncompromised host. Rev Med Virol 2003;13:155-71.
  • Kowalski RP, Karenchak LM, Romanowski EG, Gordon YJ. Evaluation of the shell vial technique for detection of ocular adenovirus. Ophthalmol 1999;106(7):1324-7.
  • Lenaerts L, De Clercq E, Naesens L. Clinical features and treatment of adenovirus infections. Rev Med Virol 2008;18:357-74.
  • Kaneko H, Maruko I, Iida T, et al. The possibility of human adenovirus detection from the conjunctiva in asymptomatic cases during nosocomial infection. Cornea 2008;5:527-30.