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Human visceral leishmaniasis (VL) caused by L. infantum and cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum have been reported in Turkey. L. infantum is also responsible for canine leishmaniasis (CanL) and it is widely common in the country. The main aim of the present study was to design a real-time PCR method based on the internal transcribed spacer 1 (ITS1) region in the diagnosis of all clinical forms of leishmaniasis in Mediterranean, and to identify the species directly from clinical samples. Totally, 315 clinical specimens, human/canine visceral (blood, bone marrow, lymph node) and cutaneous (lesion aspiration) samples, and 51 Turkish Leishmania isolates typed by isoenzymatic method were included in the study. For optimization, DNA samples of the 34 strains were amplified by conventional ITS1-PCR and then sequenced for designing the primers and probes, allowing the species identification. Following the validation with the isolates, the test was applied on clinical samples and melting temperatures were used for genotyping. A group of PCR products were further sequenced for confirmation and assigning the inter- and intraspecies heterogeneity. The diagnosis of leishmaniasis is successfully achieved by the new real-time PCR method, and the test identified 80.43% of human and canine VL samples as L.infantum and 6.52% as L.tropica; 52.46% of CL samples as L. infantum and 26.90% as L. tropica. In 13.04% of visceral and 20.62% of cutaneous samples, two peaks were observed. Hovewer, the higher peak was found to be concordant with the sequencing results in 96.96%, in terms of species identification. The real-time ITS1 PCR assay clearly identified the leishmanial species in 81.58% of all clinical samples. Genotypic variations of Leishmania parasites in Turkey within species and intraspecies were observed, and L. tropica is also found as causative agent of human and canine VL in Turkey. Author Summary: Leishmaniasis caused by Leishmania parasites are seen as cutaneous (CL) and visceral (VL) clinical forms in Turkey. Leishmania (L.) tropica and L. infantum were determined as CL agents, while L. infantum was incriminated for VL in the country. Canine leishmaniasis (CanL) is widely common throughout the country and L. infantum is the responsible agent of the disease. Related to Leishmania species diversity and different clinical forms in human and dogs in this geographical area, the identification of the parasite species prefers to be done during the time of diagnosis. Internal transcribed spacer region was chosen as the target area for developing a real-time PCR assay to use as a fast and standardized diagnostic method and species identification simultaneously. Clinical samples from parasitologically/serologically proven cases and isolates were included the study, and high positivity rate in species identification was obtained. The method can also determine the intragenomic heterogeneity in Leishmania tropica and L. infantum. The assay presents a sufficient sensitivity for fast and correct detection of leishmaniasis directly from clinical materials. L. tropica and L. infantum were found as causative agents of human CL, VL and CanL in Turkey. Knowledge about differences in the parasites is useful for future studies in Turkey. [ABSTRACT FROM AUTHOR]

Human visceral leishmaniasis (VL) caused by L. infantum and cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum have been reported in Turkey. L. infantum is also responsible for canine leishmaniasis (CanL) and it is widely common in the country. The main aim of the present study was to design a real-time PCR method based on the internal transcribed spacer 1 (ITS1) region in the diagnosis of all clinical forms of leishmaniasis in Mediterranean, and to identify the species directly from clinical samples. Totally, 315 clinical specimens, human/canine visceral (blood, bone marrow, lymph node) and cutaneous (lesion aspiration) samples, and 51 Turkish Leishmania isolates typed by isoenzymatic method were included in the study. For optimization, DNA samples of the 34 strains were amplified by conventional ITS1-PCR and then sequenced for designing the primers and probes, allowing the species identification. Following the validation with the isolates, the test was applied on clinical samples and melting temperatures were used for genotyping. A group of PCR products were further sequenced for confirmation and assigning the inter- and intraspecies heterogeneity. The diagnosis of leishmaniasis is successfully achieved by the new real-time PCR method, and the test identified 80.43% of human and canine VL samples as L.infantum and 6.52% as L.tropica; 52.46% of CL samples as L. infantum and 26.90% as L. tropica. In 13.04% of visceral and 20.62% of cutaneous samples, two peaks were observed. Hovewer, the higher peak was found to be concordant with the sequencing results in 96.96%, in terms of species identification. The real-time ITS1 PCR assay clearly identified the leishmanial species in 81.58% of all clinical samples. Genotypic variations of Leishmania parasites in Turkey within species and intraspecies were observed, and L. tropica is also found as causative agent of human and canine VL in Turkey. [ABSTRACT FROM AUTHOR]

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This study aimed to evaluate clinical signs related with canine leishmaniosis (CanL) and to determine their relation with serological results in the differential diagnosis of this disease in dog populations. A total of 253 dogs from houses and a dog shelter in KuSadasý, an endemic region for leishmaniosis, were investigated for clinical signs related to CanL. Sera collected from the study group were examined by indirect fluorescence antibody (IFA), whole ELISA and rK39 ELISA tests for the serological diagnosis. Popliteal lymph node aspiration materials were examined for the parasitological diagnosis. Clinical signs were separated into two groups as cutaneous and visceral signs and no clinical signs were observed in 51.7% (131/253) of the dogs, while 48.3% (122/253) of the dogs had at least one sign. A total of 42 dogs [29 (23.7%) in the symptomatic and 13 (9.9%) in the asymptomatic group] were diagnosed as CanL serologically and/or parasitologically. Two strains were isolated and identified as Leishmania infantum MON-1. There was no correlation between IFAT titers and the number of the clinical signs (P > 0.05; R square = 0.002). Weight loss was found to be the most common sign of CanL while epistaxis was the least common but specific sign. The seropositivity ratio of CanL is 16.6% (42/253) among dogs in the region and clinical signs were found to be helpful for suspecting CanL but at least one serological and/or parasitological method should be performed for the accurate and differential diagnosis in the dogs. Our findings also showed that the one-third of the dogs in an endemic area for CanL could be asymptomatic. [ABSTRACT FROM AUTHOR]

Copyright of Turkish Journal of Veterinary & Animal Sciences is the property of Scientific and Technical Research Council of Turkey and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Presents a study on the diagnostic value of recombinant K39 dipstick in zoonotic visceral leishmaniasis in Turkey. Identification of K39 as a repetitive immunodominant epitope in a kinesin-related protein; Comparison with other conventional diagnostic methods; Determination of the presence of parasite in humans and dogs.