Molecular methodology for the detection of the Leishmania genus in different biological samples extracted from dogs
Sá, Ana Júlia Lopes Miranda Ferreira deOlimpo, Ellen Laureanny AraújoRoyo, Vanessa de AndradeMelo Júnior, Afranio Farias deOliveira, Dario Alves deFerreira, Hanna Carolina CamposAzevedo, Alcinei MisticoMenezes, Elytania Veiga
Background: The leishmaniases are a group of parasitic diseases caused by trypanosomatids belonging to the genus Leishmania, members of the class Kinetoplastida, order Trypanosomatidae, family Trypanosomatidae. Despite innumerous wildspecies that are infected, the domestic dog is a potential reservoir of Leishmania infantum in urban areas, which expandsthe transmission pathway to humans. When infected, the dog becomes Visceral Canine Leishmaniasis (CVL), which ischaracterized by a diverse clinical picture that ranges from asymptomatic to non-specific signs, such as skin lesions, lymphadenomegaly, weight loss, splenomegaly, and/or ocular lesions, thus impairing accurate and rapid diagnosis. In Brazil, itis considered a public health problem since it is endemic in certain regions. Therefore, parasitological, serological, andmolecular methods can be used for the detection of the disease. However, the possibility of serological cross-reaction andthe occurrence of co-infection with other trypanosomatids decreases the specificity rate to below 100%, which suggeststhe use of more accurate diagnostic tools. Several molecular targets and starting samples for leishmaniasis diagnosis arealready standardized, but there is lack of data allowing the evaluation of the target, as well as which biological material ismore efficient for the molecular diagnosis of CVL. The sensitivity of PCR may vary with DNA quality, primer type, parasitemia level, and number of target copies per cell. The spleen, blood, liver, aspirate of bone marrow and lymph nodes arethe most frequently used for molecular diagnosis of CVL. The present study aimed to evaluate and compare three protocolsof the polymerase chain reaction (PCR) for the molecular diagnosis of CVL in different biological samples removed fromanimals...(AU)
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