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Visceral Leishmaniasis is considered one of the endemic parasitic diseases transferred via sand fly Phlebotomus alexandri bite that can be transferred Leishmania infantum. This study aimed to detect visceral leishmaniasis in peripheral blood for patients using IFA test and kDNA polymerase chain reaction. Eighty samples were collected in the period (September 2022 -December 2023) from patients (1-10 years of age) suspected of VL and admitted to hospitals at several governorates in Iraq. IFAT showed positive results for 65 out of 80 samples with visceral leishmaniasis by L.infantum (81%). While kDNA results showed 60 out of 80 samples with visceral leishmaniasis by L.infantum (75% )., the sensitivity and specificity of IFA were 92% and 100% comparison with PCR. Concerning the results sequencing of PCR products, there was one isolate of L.infantum that differed from the global isolates in the NCBI It was found variation sequences in position substitution including deletion, transition, and transversion in the local isolate registered with accession number OR886069.1 . Moreover, it was observed in the phylogenetic tree analysis, there were the highest and lowest similarity distances of the Iraqi isolate with other isolates that including the French isolate, and the Iranian isolate. [ABSTRACT FROM AUTHOR]