Abstract
Human adenoviruses (HAdVs) are common pathogens that can cause respiratory, gastrointestinal, urogenital, and ocular infections. They are divided into seven species containing 85 genotypes. Straightforward typing systems might help epidemiological investigations. As homologous recombination frequently shapes the evolution of HAdVs, information on a single gene is seldom sufficient to allow accurate and precise typing, and complete genome-based methods are recommended. Even so, complete genome analyses are not always easy to perform for practical reasons, and in such cases a multigene system can provide considerably more information about the strain under investigation than single-gene-based methods. Here we present a rapid, generic, multigene typing system for HAdVs based on three main deterministic regions of these viruses. Three PCR systems were used to amplify the genes encoding the DNA polymerase, the penton base hypervariable Arg-Gly-Asp-containing loop, and the hexon loop 1 (hypervariable region 1–6). Using this system, we typed 281 clinical isolates, detected members of six out of seven HAdV species (Human mastadenovirus A–F), and could also detect not only divergent strains of established types but also a new recombinant strain with a previously unpublished combination of adenovirus genomes. This strain was accepted by the Human Adenovirus Working Group as a novel genotype: HAdV-86. Seven strains that could not be typed with sufficient accuracy were also investigated using a PCR based on part of the fiber gene. By analysis of corresponding sequences of the 86 known HAdV genotypes, we determined that the proposed typing system should be able to distinguish all non-recombinant types, and with additional fiber information, all known HAdV genotypes.
Highlights
The seven species of human adenovirus (Human mastadenovirus A–G; Human adenoviruses (HAdVs)-A to HAdV-G) are divided into types, originally called serotypes
Analysis of part of the DNA polymerase gene revealed that 71.5% of the strains that gave DNA polymerase sequences belonged to species HAdV-C, but almost all other human adenovirus species were found as well, with the exception of species G
These indicated that four strains (UmU010, UmU193, UmU225, and UmU253) diverged significantly from the closest HAdV reference strain, so the four complete genomes were sequenced
Summary
We aimed to identify (1) new, divergent, distance-based types, showing low sequence identity to the closest related reference strain―earlier this would have been a new serotype, and (2) new and unpublished recombinants―new genotypes
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