DETECTION OF ADENOVIRUS DNA IN PERIPHERAL-BLOOD MONONUCLEAR-CELLS BY POLYMERASE CHAIN-REACTION ASSAY

Adenovirus can establish persistent infections which may reactivate an d cause disease in immunocompromised hosts. Lymphocytes have been post ulated to serve as a site of adenoviral persistence based upon the abi lity to isolate adenovirus from tonsils and to detect adenovirus DNA b y Southern blot hybridization in peripheral blood mononuclear cells (P BMC). To test this hypothesis, a more sensitive and specific polymeras e chain reaction (PCR) assay was developed to detect adenovirus DNA. T wo sets of nested primers were designed to conserved sequences in the adenovirus E1A and hexon genes. The E1A and hexon primers amplified DN A from representative adenoviral serotypes in all six adenoviral group s (A-F). Both primers detected a single copy of the adenovirus type 2 genome but were less sensitive for the group B type 35. None of 33 PBM C specimens from healthy adults and only one of 40 pediatric samples w as positive (at a low level) for adenovirus DNA by nested PCR assay. I n comparison, PBMC from two children with fatal adenoviral infection w ere both strongly positive for adenovirus DNA. It is concluded that, i n contrast to a previous study, PBMC are not a common site of persiste nt group C adenoviral infection. In addition, assay of PBMC by the ade novirus-specific PCR may help detect early invasive disease and warran ts further evaluation. (C) 1997 Wiley-Liss, Inc.