Genotyping of clinically relevant human adenoviruses by array-in-well hybridization assay.
A robust oligonucleotide array-in-well hybridization assay using novel up-converting phosphor reporter technology was applied for genotyping clinically relevant human adenovirus types. A total of 231 adenovirus-positive respiratory, ocular swab, stool and other specimens from 219 patients collected between April 2010 and April 2011 were included in the study. After a real-time PCR amplification targeting the adenovirus hexon gene, the array-in-well assay identified the presence of B03 (n = 122; 57.5% of patients), E04 (29; 13.7%), C02 (21; 9.9%), D37 (14; 6.6%), C01 (12; 5.7%), C05 (5; 2.4%), D19 (4; 1.9%), C06 (2; 0.9%), D08 (1; 0.5%), A31 (1; 0.5%) and F41 (1; 0.5%) genotypes among the clinical sample panel. The typing result was obtained for all specimens that could be amplified (n = 223; 97%), and specificity of the typing was confirmed by sequencing specimens representing each of the different genotypes. No hybridization signal was obtained in adenovirus-negative specimens or specimens with other viruses (n = 30). The array-in-well hybridization assay has great potential as a rapid and multiplex platform for the typing of clinically relevant human adenovirus genotypes in different specimen types.
Department of Virology, University of Turku, Turku, Finland. minna.yliharsila@utu.fi
M
M
Ylihärsilä
E
E
Harju
R
R
Arppe
L
L
Hattara
J
J
Hölsä
P
P
Saviranta
T
T
Soukka
M
M
Waris
D000258:Adenovirus Infections, Human / Q000175:diagnosis / Q000453:epidemiology; D000260:Adenoviruses, Human / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D000293:Adolescent; D000328:Adult; D000368:Aged; D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D005260:Female; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction; D012621:Seasons; D055815:Young Adult
D036022:Capsid Proteins; C018315:hexon capsid protein, Adenovirus