| The high prevalence of Leishmania infection was reported in dogs as the main reservoir of CanL in many locations
in the old world. Detection and firmly identification of Leishmania species in asymptomatic dogs by reliable
method was considered and employed. Non-invasive and non-anesthetized blood sampling in asymptomatic dogs
was conducted. Nested, conventional and real-time PCR with HRM technique was performed targeting ITS-rDNA
gene. 88 asymptomatic dogs were sampled from three CanL endemic provinces of Iran in 2018–2019. 23 blood
samples were Leishmania positive. L. major, L. tropica and L. infantum were accurately identified for the first time
with HRM targeting ITS2-microsatellite. Three samples were mixed infections. CLC software TM predictions for
microsatellite ITS-rDNA were 86.93
◦
C: L. major, 85.76
◦
C: L. tropica and 86.04
◦
C: L. infantum. Standard strains
of Leishmania species were accurately separated with almost one to 2
◦
C deference (L. major: 86.61
◦
C,
L. infantum: 85.41
◦
C, L. tropica: 84.82
◦
C). Each HRM curve represents one species in a sample for helping
accurate identification of Leishmania species and even mixed infection when two curves are present. Detecting
parasites at primary stages in asymptomatic cases is essential using Real-time HRM. As same as mammalian
Leishmania in rodents which is present at early stages and non-pathogenesis, only L. major would exist and other
Leishmania disappears. This can conclude also for L. major, L. infantum and L. tropica in dogs. The role of L. major
existence in canine blood should be investigated more. |