Adenovirus polymerase chain reaction assay for rapid diagnosis of conjunctivitis.
OBJECTIVE
To evaluate newly designed primers in a polymerase chain reaction (PCR) for the detection of adenovirus DNA in conjunctival swabs.


METHODS
Oligonucleotides were derived from the adenovirus hexon gene and modified such that a maximum of only two mismatches occurred with adenovirus types 2 through 5, 7, and 16. Specificity was determined against adenovirus types 2 through 4, 7, 8 through 11, 14, 19, 37, 40, and 41 and from non-adenoviral DNA and the sensitivity by PCR amplification of purified adenovirus type 2 DNA. The assay was compared retrospectively with cell culture and a PCR with different primers on 59 stored conjunctival swab samples. The new PCR also was used prospectively in comparison with cell culture on 2743 conjunctival swabs.


RESULTS
The 140-bp product was amplified from all the adenovirus serotypes tested except types 40 and 41, which have not been isolated from the eye. There were no amplified products from the non-adenoviral DNA tested. With adenovirus type 2 DNA, despite two deliberate mismatches, 40 copies of the target were detectable after PCR and ethidium bromide-staining. In the retrospective study, 51 of 55 (92.7%) were positive by this new PCR compared with 42 of 55 (76.4%) by the older PCR and 40 of 55 (72.7%) by cell culture. In the prospective study, the new PCR detected 386 of 415 (93%) adenovirus-positive specimens compared with 248 of 415 (59.8%) by cell culture. Of 167 specimens positive for herpes simplex virus by cell culture, none were positive by the adenovirus PCR.


CONCLUSIONS
PCR with the newly designed primers shows a much increased sensitivity over cell culture and previous PCRs for the detection of adenoviruses in conjunctival swabs.
Cooper
R J
RJ
Department of Pathologic Sciences, University of Manchester, England.
Yeo
A C
AC
Bailey
A S
AS
Tullo
A B
AB
D000258:Adenovirus Infections, Human / Q000175:diagnosis* / Q000821:virology; D000260:Adenoviruses, Human / Q000235:genetics* / Q000302:isolation & purification; D002478:Cells, Cultured; D003228:Conjunctiva / Q000166:cytology / Q000821:virology; D003236:Conjunctivitis, Viral / Q000175:diagnosis* / Q000821:virology; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D011446:Prospective Studies; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral