Abstract: Herpes simplex keratitis is one of the important causes of blindness and its early diagnosis is essential to the treatment of the respective patients accordingly. The aim of this study was to diagnose and analyze quantitatively the herpes simplex DNA in patients with suspected Herpes keratitis using TaqMan real-time PCR method. Corneal swabs from HSV keratitis suspected patients were collected from September 2005 to December 2009. Upon DNA extraction, the samples were analyzed by quantitative TaqMan real-time PCR assay. In this method, a set of primers amplified a common sequence of HSV-1 and HSV-2 glycoprotein B gene. The copy number of unknown samples were expressed via standard curve drawn with known amount of amplified cloned plasmid containing target sequences. Of the 296 samples, 178 (60.1%) belonged to males and 118 (39.9%) to females. The HSV DNA was detected in 111 (37.5%) of the cases by PCR, consisting of 69 males and 42 females. The drawn standard curve was linear in 10 to 106 copies of virus (R2 = 0.982). The ranges of the HSV DNA copy numbers in the collected samples were detected from 2.7x103 to 6.1x105 copies/samples. Present results suggest that Herpes simplex keratitis remains as an epidemiologically important eye disease and improvement in the monitoring and detecting of ocular herpetic disease by quantitative PCR method is informative and helpful to the clinical diagnosis, decision for treatment and follow up.