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.
Kinchington
P R
PR
Charles T. Campbell Laboratory for Infectious Diseases, Department of Ophthalmology, University of Pittsburgh, Pennsylvania 15213.
Turse
S E
SE
Kowalski
R P
RP
Gordon
Y J
YJ
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