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Journal of Clinical Microbiology, March 2007, p. 958-967, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01603-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Quantitative Detection and Rapid Identification of Human Adenoviruses{triangledown}

Rika Miura-Ochiai,1 Yasushi Shimada,1 Tsunetada Konno,1 Shudo Yamazaki,1 Koki Aoki,2 Shigeaki Ohno,3 Eitaro Suzuki,4 and Hiroaki Ishiko1*

Research and Development Department, Mitsubishi Kagaku Bio-Clinical Laboratories, Inc., Tokyo 174-8555, Japan,1 Department of Ophthalmology, Yokohama City University School of Medicine, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan,2 Department of Ophthalmology and Visual Sciences, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido 060-8638, Japan,3 Suzuki Pediatric Clinic, Ube, Yamaguchi 755-0151, Japan4

Received 3 August 2006/ Returned for modification 24 September 2006/ Accepted 23 December 2006

We have established a method of quantitative detection and rapid identification of human adenoviruses (hAdVs). Using LightCycler PCR with a primer set, we were able to amplify 554 bp of the hexon gene from each of 51 prototype strains of hAdVs. The sensitivity of LightCycler PCR was 10 copies of hAdV DNA/reaction. When LightCycler PCR was performed using a set of primers, hAdV was positive for 74.4% (99 of 133) of conjunctivitis patients and for 27.3% (81 of 297) of respiratory infection patients. We also attempted to measure hAdV in the potentially contaminated eye drops used by patients, detecting 5.4 x 102 to 1.6 x 106 copies/ml of hAdV. We determined the 350-bp nucleotide sequence of the amplified hexon gene and compared it with the sequences of the 51 prototype strains. Phylogenetic analysis based on 350 bp of the hexon gene identified 99 positive conjunctival swabs as 24 cases of AdV type 3 (AdV-3), 14 cases of AdV-4, 1 case of AdV-8, 19 cases of AdV-19a, and 41 cases of AdV-37. The 81 sequences from pharyngeal or nasal mucus swabs were identified as 29 cases of AdV-2, 18 cases of AdV-1, 18 cases of AdV-5, 12 cases of AdV-4, 2 cases of AdV-37, 1 case of AdV-3, and 1 case of AdV-6. LightCycler PCR followed by phylogenetic analysis provides an effective tool for the rapid identification of hAdVs and for studying molecular epidemiology.

* Corresponding author. Mailing address: Research and Development Department, Mitsubishi Kagaku Bio-Clinical Laboratories, Inc., Shimura 3-30-1, Itabashi-ku, Tokyo 174-8555, Japan. Phone: 81 3 5994 2431. Fax: 81 3 5994 2972. E-mail: Ishiko{at}nm.mbcL.co.jp.

{triangledown} Published ahead of print on 17 January 2007.

Journal of Clinical Microbiology, March 2007, p. 958-967, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01603-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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