MULTIPLEX POLYMERASE CHAIN-REACTION FOR ADENOVIRUS AND HERPES-SIMPLEXVIRUS IN EYE SWABS

Adenoviruses and herpes simplex virus (HSV) can cause clinically indis tinguishable episodes of acute eye disease. Adenovirus infection is as sociated with nosocomial outbreaks and HSV may result in episodes of r ecurrent ocular inflammation. In a comparison of multiplex PCR for the two viral DNAs and virus isolation in cell culture, identical results were obtained for 18 of 20 specimens (positive for adenovirus in 5, H SV in 5, and negative in 8). One specimen was falsely negative for eac h viral DNA. Inclusion of human beta-globin primers in the adenovirus- HSV reaction was precluded by a consequential 10-100-fold reduction in sensitivity for the two viral targets and by the failure of beta-glob in DNA amplification at the annealing temperature (45 degrees C) requi red to ensure detection of adenoviruses of serotypes 7 and 11 with the selected adenovirus primers. A single-target beta-globin PCR gave pos itive results with 19 of the 20 specimens prepared by treatment with p roteinase K lysis buffer, indicating the effectiveness of this simple DNA extraction procedure. Nonetheless, the availability of effective a ntiviral therapy for HSV made monitoring for extraction failure using human primers crucial to avoid false-negative results for HSV DNA. Ade novirus-HSV PCR has considerable potential for the rapid diagnosis of viral eye disease particularly if beta-globin primers can be included in the reaction.