title: Detection of adenovirus DNA in peripheral blood mononuclear cells by polymerase chain reaction assay.
abstract: Adenovirus can establish persistent infections which may reactivate and cause disease in immunocompromised hosts. Lymphocytes have been postulated to serve as a site of adenoviral persistence based upon the ability to isolate adenovirus from tonsils and to detect adenovirus DNA by Southern blot hybridization in peripheral blood mononuclear cells (PBMC). To test this hypothesis, a more sensitive and specific polymerase chain reaction (PCR) assay was developed to detect adenovirus DNA. Two sets of nested primers were designed to conserved sequences in the adenovirus E1A and hexon genes. The E1A and hexon primers amplified DNA from representative adenoviral serotypes in all six adenoviral groups (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 PBMC specimens from healthy adults and only one of 40 pediatric samples was positive (at a low level) for adenovirus DNA by nested PCR assay. In comparison, PBMC from two children with fatal adenoviral infection were both strongly positive for adenovirus DNA. It is concluded that, in contrast to a previous study, PBMC are not a common site of persistent group C adenoviral infection. In addition, assay of PBMC by the adenovirus-specific PCR may help detect early invasive disease and warrants further evaluation.
authors.affiliation: Department of Medicine, Medical College of Wisconsin, Milwaukee, USA.
authors.forename: P
authors.initials: P
authors.lastname: Flomenberg
authors.forename: E
authors.initials: E
authors.lastname: Gutierrez
authors.forename: V
authors.initials: V
authors.lastname: Piaskowski
authors.forename: J T
authors.initials: JT
authors.lastname: Casper
mesh_terms: D000257:Adenoviridae Infections / Q000175:diagnosis / Q000821:virology; D017500:Adenovirus E1A Proteins / Q000235:genetics; D000260:Adenoviruses, Human / Q000145:classification / Q000302:isolation & purification*; D000328:Adult; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000235:genetics; D036022:Capsid Proteins; D002648:Child; D017124:Conserved Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood*; D006801:Humans; D008214:Lymphocytes / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012016:Reference Values; D012680:Sensitivity and Specificity; D012703:Serotyping
chemical_list: D017500:Adenovirus E1A Proteins; D000956:Antigens, Viral; D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; C018315:hexon capsid protein, Adenovirus