Background: Cutaneous leishmaniasis (CL) remains a serious public health concern in some parts of Iraq. The aims of this study to report of CL in some parts of Iraq, by different parasitological, cultural, and molecular methods and evaluate sex differences among infected patients. This is the first study conducted to characterize Leishmania species causing CL among Iraqi patients using the sequence analysis of Internal Transcribed Spacer1 (ITS1). Methods: A total of 700 cases of suspected CL who were referred to the Iraqi's clinics and health centers and they checked for Leishmania amastigote using Giemsa-stained , however the NovyMacneal Nicolle (NNN) culture led to the growth of promastigotes in all samples, then the samples were examined using PCR and RFLP-PCR methods. Results: The present study indicated that the prevalence of CL in eight Iraqi provinces as follow: Diwaniya 88(15.1%) , Wasit 85(14.5%),Najaf 79(13.6%), Thi-Qar 76(13.1%), Basrah 67(11.5%), Baghdad 65(11.2%), Diyala 63(10.8%) and Salah-Adin province which recorded the lower infection 60(10.3%), and it appeared 83.3% by using Giemsa-smeared in eight Iraqi provinces. The highest infection (100 %) appeared using PCR while the lowest infection (68%) appeared by culture on NNN media. The present study was revealed that the highest infection (60 %) caused by L.major than L.tropica (40 %) . Our results showed that 368 (52.6 %) of CL patients were had single lesion and 215 (30.7 %) had multiple lesions, and the ulcerative wet type lesions were present in 49.6% , while the nodule dry type lesions were present in 33.7 %. The overall prevalence of cutaneous leishmaniasis in the study area was very high (83.3%) having a statistical significant association with sex; males are more prone (56.4%) to Cutaneous Leishmaniasis (CL) as compared to females(43.6%). Conclusions: A clear and reliable bias toward males exists in some tropical diseases, such as leishmaniasis. Cutaneous leishmaniasis is a major health problem in Iraq and CL caused by many countries including Iraq. Health authorities should be aware of the fact that war and terrorist activities induce expansion of the disease and increase incidence rate in the situation that access to medical treatment is not easy especially in poor conditions of leishmaniasis endemic areas.