A new set of PCR assays for the identification of multiple human adenovirus species in environmental samples.
OBJECTIVE
To assess human adenovirus (HAdV) diversity in environmental samples based on sequence comparisons of hexon gene fragments amplified using newly designed HAdV-specific polymerase chain reaction (PCR) assays.


RESULTS
Six PCR primer sets were designed based on 56 aligned hexon sequences from NCBI GenBank to amplify different hexon gene sections (241-349 bp) of the six HAdV species. The amplified hexon genes from wastewater samples were cloned, sequenced, and compared with those in publicly accessible databases (i.e. NCBI GenBank) by using the Blast program. A total of 46 analysed positive clones were affiliated to five HAdV serotypes, i.e. 1, 2, 12, 31 and 41. Similarities between the cloned and database hexon sequences ranged from 95.9 to 100% (with an average of 98.1 +/- 1.0%).


CONCLUSIONS
The designed primers showed higher amplification efficiencies when compared with the existing assays. Using the new assays, HAdV species A, C, and F (serotypes 1, 2, 12, 31 and 41 in particular) were identified in the studied municipal wastewater.


CONCLUSIONS
The six PCR primer sets developed in this study can be used to efficiently amplify hexon gene fragments in HAdV. Multiple HAdV serotypes identified in the municipal wastewater provide new information about HAdV diversity in environmental samples.
Kuo
D H-W
DH
Department of Civil and Environmental Engineering, Michigan State University, East Lansing, MI 48824, USA.
Simmons
F
F
Xagoraraki
I
I
D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D036022:Capsid Proteins / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods; D005814:Genes, Viral; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D012703:Serotyping; D012722:Sewage / Q000382:microbiology*
D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; D012722:Sewage; C018315:hexon capsid protein, Adenovirus