Adenovirus and Herpes simplex virus (HSV) are common viral causes of keratoconjunctivitis (KC). A rapid laboratory diagnosis is often very beneficial. The aim of this study was to show the prevalence of Adenovirus and HSV in patients diagnosed as KC and to evaluate the efficacy of rapid direct antigen detection of Adenovirus and HSV-1 by direct immunofluoresence assay (DIF) compared with Vero cell line culture for diagnosis of viral KC. Fifty eight patients clinically diagnosed as viral KC were included in this study; forty patients with follicular conjunctivitis and eighteen patients with keratitis. Conjunctival and corneal scraps were tested by DIF for Adenovirus and HSV compared to viral isolation which was performed on Vero cell line. Adenovirus was detected in 45% and 5.6% of conjunctivitis and keratitis patients respectively while HSV was detected in 27.5% and 38.9% respectively. Adenovirus was detected in 32.8% of patients by DIF while 13.8% by cell culture. Positive predictive value (PPV), Negative predictive value (NPV),Sensitivity, specificity, and efficiency of DIF for Adenovirus were 42.1%, 100% 100%, 78%, and 81% respectively as compared to cell culture .HSV was detected in 27.6% by DIF while 8.6% by cell culture. Sensitivity, specificity, PPV, NPV and efficiency of DIF for HSV were 60%, 75.5%, 18.8%, 95.2% and 74.1% respectively as compared to cell culture. From this study we conclude that Adenovirus was a common pathogen in conjunctivitis and HSV was a common pathogen of KC. IFA is rapid and simple test and can be used as primary screening tool. The combination of cell culture and DIF establishes the best set of methods of diagnosis of clinically suspected infected cases of viral KC.