DETECTION AND TYPING OF SUBGROUP-F ADENOVIRUSES USING THE POLYMERASE CHAIN-REACTION

Citation
Ct. Tiemessen et Mj. Nel, DETECTION AND TYPING OF SUBGROUP-F ADENOVIRUSES USING THE POLYMERASE CHAIN-REACTION, Journal of virological methods, 59(1-2), 1996, pp. 73-82
Citations number
39
Categorie Soggetti
Virology,"Biochemical Research Methods","Biothechnology & Applied Migrobiology
ISSN journal
01660934
Volume
59
Issue
1-2
Year of publication
1996
Pages
73 - 82
Database
ISI
SICI code
0166-0934(1996)59:1-2<73:DATOSA>2.0.ZU;2-I
Abstract
A DNA amplification test was developed for the sensitive detection of the diarrhoea-associated subgroup F adenoviruses in clinical specimens . The test was made highly specific for serotypes 40 and 41 by using a region of the genome (the long-fiber gene) which is not significantly homologous to other human adenoviruses, but which is highly conserved between Ad40 and Ad41. A positive subgroup F adenovirus diagnosis was characterized by the presence of an amplification product of 152 base pairs, which could be digested into products of predictable length by restriction enzymes XbaI and SpeI. The viruses were typed as either A d40 or Ad41 by digestion of the amplification product with a restricti on enzyme which digested only Ad40 DNA. The specificity of the test wa s assessed using DNA from other adenoviruses, from human and simian ce lls, and from bacteria commonly found in the human intestine. There wa s a strong correlation between results of typing obtained with PCR and restriction enzyme typing of Ad40 and Ad41, and also positivity using subgroup F specific probes in dot blot hybridizations.