title: PCR and restriction endonuclease analysis for rapid identification of human adenovirus subgenera.
abstract: Subgenus identification of adenoviruses is of clinical importance and is as informative as identification by serotype in most clinical situations. A PCR-based identification of adenovirus subgenera A, B, C, D, E, and F and sometimes serotypes is described. The PCR uses nonnested primer pair ADRJC1-ADRJC2, which targets a highly conserved region of the adenovirus hexon gene, has a sensitivity of 10 to 40 copies of adenovirus type 2 (Ad2) DNA, and generates 140-bp PCR products from adenovirus serotypes representative of all the subgroups. The PCR products of all subgroups can be differentiated on the basis of the restriction fragment patterns produced by a total of five restriction endonucleases. In addition, serotypes Ad40 and Ad41 (subgroup F) and important serotypes of subgroup D (Ad8, Ad10, Ad19, and Ad37) can easily be differentiated, but serotypes within subgroups B and C cannot. The method was assessed by blind subgenus identification of 56 miscellaneous clinical isolates of adenoviruses. The identities of these isolates at the subgenus level by the PCR correlated 91% (51 of 56) with the results of serotyping by the neutralization test, and 9% (5 of 56) of clinical isolates produced discordant results.
authors.affiliation: School of Medicine, The University of Manchester, Manchester M13 9WL, United Kingdom.
authors.forename: E M
authors.initials: EM
authors.lastname: Elnifro
authors.forename: R J
authors.initials: RJ
authors.lastname: Cooper
authors.forename: P E
authors.initials: PE
authors.lastname: Klapper
authors.forename: A S
authors.initials: AS
authors.lastname: Bailey
mesh_terms: D000258:Adenovirus Infections, Human / Q000821:virology; D000260:Adenoviruses, Human / Q000145:classification* / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000235:genetics; D036022:Capsid Proteins; D017124:Conserved Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
chemical_list: D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; C018315:hexon capsid protein, Adenovirus