Quantitative detection and rapid identification of human adenoviruses.
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.
Miura-Ochiai
Rika
R
Research and Development Department, Mitsubishi Kagaku Bio-Clinical Laboratories, Inc., Shimura 3-30-1, Itabashi-ku, Tokyo 174-8555, Japan.
Shimada
Yasushi
Y
Konno
Tsunetada
T
Yamazaki
Shudo
S
Aoki
Koki
K
Ohno
Shigeaki
S
Suzuki
Eitaro
E
Ishiko
Hiroaki
H
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
D036022:Capsid Proteins; D004279:DNA, Viral; C018315:hexon capsid protein, Adenovirus