USE OF POLYMERASE CHAIN AMPLIFICATION REACTION FOR THE DETECTION OF ADENOVIRUSES IN OCULAR SWAB SPECIMENS

Purpose. To evaluate the application of polymerase chain reaction (PCR ) methodology as a potential diagnostic tool for the detection of aden ovirus DNA in ocular swab samples. Methods. Oligonucleotides derived f rom the adenovirus hexon gene were used to amplify a 306-base pair (bp ) product by PCR. Radiolabeled oligonucleotides derived from sequences within the amplified product were used as specific probes. Specificit y was determined against DNA of 13 adenovirus serotypes (types 1 to 11 , inclusive, and types 19 and 37) and from nonadenoviral DNAs. Limits of detection were determined by PCR amplification of known amounts of purified adenovirus serotype 2 DNA. The assay was tested on 107 ocular swab samples and correlated to results obtained from tissue culture a nd a commercial immunoassay (Adenoclone). Results. The 306-bp PCR prod uct was amplified from all adenovirus serotypes tested, but not from n egative control DNAs. As little as 15 fg of adenovirus type 2 DNA coul d be detected by PCR and ethidium bromide stain. Using a simplified sa mple preparation procedure, 46 of 58 adenovirus culture-positive but A denoclone-negative swabs were positive by PCR (79% sensitivity). All ( 11 of 11) Adenoclone-positive clinical eye swabs tested were positive by PCR (100% sensitivity). Only 1 of 38 nonadenoviral ocular swab samp les was positive by PCR (97% specificity). Conclusions. PCR appeared t o be highly suitable for the diagnosis of adenovirus in ocular swabs, offering important improvements in speed over tissue culture isolation and in sensitivity over immunoassay.