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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]

• Better performance was obtained compared to more commonly used antigens. • Dyn-1 showed excellent ability to identify asymptomatic dogs (100% sensitivity). • Overall, the ability of rDyn-1 to identify dogs with CanL was 97% (sensitivity). • rDyn-1-based ELISA assays could be advantageous over classic diagnostic tests. Visceral leishmaniasis (VL) is a fatal manifestation of an infection caused by intracellular protozoa of the Leishmania genus. In New World countries, VL is classified as a zoonotic disease with domestic dogs acting as its main reservoir. Asymptomatic dogs are as competent to transmit Leishmania to the vectors as symptomatic dogs, however current diagnostic tests are limited and present low sensitivity for this important group. The development of accurate tests is fundamental to the early diagnosis, treatment, and control of canine leishmaniasis. In this study, we investigated the use of a recombinant protein (dynamin-1-like protein, Dyn-1) from L. infantum, as a potential target antigen for leishmaniasis serodiagnosis in both symptomatic and asymptomatic dogs. The antigenic performance of the protein was evaluated by means of ELISA assays using sera from symptomatic (n = 25), asymptomatic (n = 34) and non-infected dogs (n = 36) using ELISA. In addition, sera from dogs experimentally infected with Trypanosoma cruzi (n = 49) and naturally infected with Babesia sp. (n = 8) were tested to evaluate possible cross-reactivity. A crude soluble antigen (CSA) of Leishmania was used as an antigen control and K39 and K26 were used as reference antigens because they are already widely used in commercial tests. rDyn-1-based assay showed the highest sensitivity (97%) compared to the antigens K39 (88%), K26 (86%) and crude extract (95%). The highest specificity among the tests was also obtained with the protein rDyn-1 (94%), compared with the other antigens K39 (81%), K26 (87%), and crude extract (77%). This study showed that the rDyn-1 ELISA assay was able to identify 100% of asymptomatic dogs, establishing its potential as a target for the diagnosis of canine leishmaniasis. [ABSTRACT FROM AUTHOR]