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Journal of Clinical Microbiology, June 2000, p. 2055-2061, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

PCR and Restriction Endonuclease Analysis for Rapid Identification of Human Adenovirus Subgenera

Elfath M. Elnifro,1 Robert J. Cooper,1,* Paul E. Klapper,1,2 and Andrew S. Bailey2

School of Medicine, The University of Manchester,1 and Clinical Virology, Central Manchester Healthcare Trust,2 Manchester M13 9WL, United Kingdom

Received 10 November 1999/Returned for modification 31 January 2000/Accepted 10 March 2000

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.

* Corresponding author. Mailing address: University Virology, 3rd Floor, Clinical Sciences Building, Central Manchester Healthcare Trust, Oxford Road, Manchester M13 9WL, United Kingdom. Phone: 44 (0)161-276-8844. Fax: 44 (0)161-276-8840. E-mail: Bob.Cooper{at}man.ac.uk.

Journal of Clinical Microbiology, June 2000, p. 2055-2061, Vol. 38, No. 6
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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