PCR and restriction endonuclease analysis for rapid identification of human adenovirus subgenera

Robert Cooper, Elfath M. Elnifro, Robert J. Cooper, Paul E. Klapper, Andrew S. Bailey

    Research output: Contribution to journalArticlepeer-review

    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.
    Original languageEnglish
    Pages (from-to)2055-2061
    Number of pages6
    JournalJournal of clinical microbiology
    Volume38
    Issue number6
    Publication statusPublished - 2000

    Keywords

    • virology: Adenovirus Infections, Human
    • classification: Adenoviruses, Human
    • Base Sequence
    • genetics: Capsid
    • Capsid Proteins
    • Comparative Study
    • Conserved Sequence
    • DNA Primers
    • isolation & purification: DNA, Viral
    • Humans
    • Molecular Sequence Data
    • methods: Polymerase Chain Reaction
    • Polymorphism, Restriction Fragment Length
    • Sensitivity and Specificity
    • Sequence Analysis, DNA
    • Sequence Homology, Nucleic Acid

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