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 P CR-based identification of adenovirus subgenera A, B, C, D, E, and F and so metimes serotypes is described. The PCR uses nonnested primer pair ADRJC1-A DRJC2, 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 a ll the subgroups. The PCR products of all subgroups can be differentiated o n the basis of the restriction fragment patterns produced by a total of fiv e restriction endonucleases. In addition, serotypes Ad40 and Ad41 (subgroup F) and important serotypes of subgroup D (Ad8, Ad10, Ad19, and Ad37) can e asily be differentiated, but serotypes within subgroups B and C cannot. The method was assessed by blind subgenus identification of 56 miscellaneous c linical isolates of adenoviruses. The identities of these isolates at the s ubgenus level by the PCR correlated 91% (51 of 56) with the results of sero typing by the neutralization test, and 9% (5 of 56) of clinical isolates pr oduced discordant results.