Evaluation of polymerase chain reaction for the detection of adenoviruses in conjunctival swab specimens using degenerate primers in comparison with direct immunofluorescence.
OBJECTIVE
To investigate a rapid and sensitive polymerase chain reaction (PCR)-based assay for the detection of adenoviral infections in conjunctival swabs.


METHODS
Degenerate primers derived from the adenoviral hexon gene were designed for amplification of a 302-bp product by PCR. In 15 ocular swab specimens, using a simple sample preparation method, PCR was compared with a commercially available direct immunofluorescence test.


RESULTS
Fourteen samples of patients with clinically diagnosed adenoviral keratoconjunctivitis, when screened by direct immunofluorescence, were positive for adenoviral protein. In all samples positive by immunofluorescence, the 302-bp product was amplifiable. The sample negative by direct immunofluorescence was positive using PCR.


CONCLUSIONS
PCR using degenerate primers proved to have several advantages over current diagnostic techniques. It offers considerable improvement in sensitivity over immunoassays and speed over tissue culture isolation and is a highly potential tool for the diagnosis of adenoviral ocular infections.
Weitgasser
Ursula
U
Department of Ophthalmology, Karl Franzens University of Graz, Graz, Austria.
Haller
Eva Maria
EM
El-Shabrawi
Yosuf
Y
D000257:Adenoviridae Infections / Q000175:diagnosis; D000260:Adenoviruses, Human / Q000235:genetics / Q000302:isolation & purification*; D003228:Conjunctiva / Q000821:virology*; D003236:Conjunctivitis, Viral / Q000175:diagnosis; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D007634:Keratitis / Q000821:virology; D016133:Polymerase Chain Reaction / Q000592:standards*
D017931:DNA Primers; D004279:DNA, Viral