title: Quantitative detection and rapid identification of human adenoviruses.
abstract: 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 10(2) to 1.6 x 10(6) 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.
authors.affiliation: Research and Development Department, Mitsubishi Kagaku Bio-Clinical Laboratories, Inc., Shimura 3-30-1, Itabashi-ku, Tokyo 174-8555, Japan.
authors.forename: Rika
authors.initials: R
authors.lastname: Miura-Ochiai
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shimada
authors.forename: Tsunetada
authors.initials: T
authors.lastname: Konno
authors.forename: Shudo
authors.initials: S
authors.lastname: Yamazaki
authors.forename: Koki
authors.initials: K
authors.lastname: Aoki
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Ohno
authors.forename: Eitaro
authors.initials: E
authors.lastname: Suzuki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Ishiko
mesh_terms: D000258:Adenovirus Infections, Human / Q000175:diagnosis* / Q000821:virology*; D000260:Adenoviruses, Human / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D036022:Capsid Proteins / Q000235:genetics*; D002460:Cell Line; D003236:Conjunctivitis, Viral / Q000175:diagnosis / Q000821:virology; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000821:virology; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013997:Time Factors
chemical_list: D036022:Capsid Proteins; D004279:DNA, Viral; C018315:hexon capsid protein, Adenovirus