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Adenoviral conjunctivitis is a widespread worldwide problem in Iraq, as many adenoviral conjunctivitis patients are diagnosed each year. There are, however, few studies into pathogenic adenovirus genotypes causing conjunctivitis and a particular serotype causing this disease in mid effurates/Iraq. The goal of this research is the genotyping of adenovirus that causes conjunctivitis. Out of 150 eyeswab samples, VTM transport was then screened for the presence of HAdVs, from December 2018 to July 2019,from patients with viral conjunctivitis symptoms in Babylon, Kerbela, and Najef, Iraq respectively. Molecular detection by CYBR green qPCR amplification and nucleotide sequence outlined by viral DNA extractions. Of the 150 samples, 115 HAdVs samples (76.66%) were detected most frequency among females 56% (63/115%). Of the 30 HAdVs samples, 30 Hexon partial genes could be analyzed. The study of the sequence DNA and building phylogenetic tree, based on the hexon gene, indicate two different forms of HAdVs serotype 4 in Babylon(33.33%) and serotype 7 in kerbala, Najaf respectively(66.67%) associated with HAdVs with the significant difference 0.001. Gender distribution are presented that male 16(53.33%) and female was 14 (46.67%) that male is more excessive than female in mideffurate conjunctivitis patients / Iraq. [ABSTRACT FROM AUTHOR]