Use of polymerase chain amplification reaction for the detection of adenoviruses in ocular swab specimens.
OBJECTIVE
To evaluate the application of polymerase chain reaction (PCR) methodology as a potential diagnostic tool for the detection of adenovirus DNA in ocular swab samples.


METHODS
Oligonucleotides derived from 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. Specificity 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 and a commercial immunoassay (Adenoclone).


RESULTS
The 306-bp PCR product was amplified from all adenovirus serotypes tested, but not from negative control DNAs. As little as 15 fg of adenovirus type 2 DNA could be detected by PCR and ethidium bromide stain. Using a simplified sample preparation procedure, 46 of 58 adenovirus culture-positive but Adenoclone-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 samples was positive by PCR (97% specificity).


CONCLUSIONS
PCR appeared to 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.
Charles T. Campbell Laboratory for Infectious Diseases, Department of Ophthalmology, University of Pittsburgh, Pennsylvania 15213.
P R
PR
Kinchington
S E
SE
Turse
R P
RP
Kowalski
Y J
YJ
Gordon
D000208:Acute Disease; D000258:Adenovirus Infections, Human / Q000175:diagnosis / Q000821:virology*; D000260:Adenoviruses, Human / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000032:analysis; D001483:Base Sequence; D003236:Conjunctivitis, Viral / Q000175:diagnosis / Q000821:virology*; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D007118:Immunoassay; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D017931:DNA Primers; D004279:DNA, Viral