PCR and restriction endonuclease analysis for rapid identification of human adenovirus subgenera.
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.
Elnifro
E M
EM
School of Medicine, The University of Manchester, Manchester M13 9WL, United Kingdom.
Cooper
R J
RJ
Klapper
P E
PE
Bailey
A S
AS
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
D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; C018315:hexon capsid protein, Adenovirus