Herpes simplex virus (HSV) is a common human pathogen that causes severe infections in newborns and immunocompromised patients. Conjunctivitis or corneal epithelial keratitis is caused by HSV type 1 all over the world and at all times of the year. The present study was aimed at detecting HSV in patients suffering from conjunctivitis. One hundred and ten (110) clinical samples (90 patients and 20 controls, both males and females) of eye conjunctiva swabs were collected from patients of different ages. The samples were analyzed using qPCR and ELISA techniques. The qPCR results revealed that HSV was present in 47 (52.2%) of the 90 patients who were infected. Of these patients, 25 (48.0%) were males and 22 (57.8%) were females, indicating that females are more susceptible to infection. According to the results by age group, patients over 50 years old had a higher rate (81.8%), making young adults more susceptible to infection. The prevalence of HSV-positive results by ELISA was observed in 29 (32.3%) individuals, including 16 (30.7%) males and 13 (34.2%) females, indicating that females are more susceptible to infection. Adults over the age of 50 (54.5%) are more susceptible to infection, according to the age group’s HSV-positive outcomes. HSV type 1 infection is highly prevalent among Iraqi conjunctivitis patients, with a statistically significant difference when compared to controls, based on the two techniques. The findings of this study indicate that qPCR is more accurate and reliable than the ELISA technique for detecting HSV type 1.