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      Performance of IFAT, ELISA, direct parasitological examination and PCR on lymph node aspirates for canine visceral leishmaniasis diagnosis

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          Abstract

          Canine visceral leishmaniasis (CVL) is endemic in numerous Brazilian regions. The greatest difficulty in controlling the disease is the diagnostic limitation. In the present study, the most common tests employed for visceral leishmaniasis diagnosis were compared: immunofluorescence antibody test (IFAT), immunoenzymatic assay (ELISA), direct parasitological examination and polymerase chain reaction (PCR). Samples of lymph node aspirates and blood were collected from 100 dogs that lived in an endemic area (Bauru city, São Paulo state) and from 100 negative controls from a non-endemic area (Botucatu city, São Paulo state). Specificity of both IFAT and PCR was 100% whereas ELISA was 99%. Sensitivities were 97.77, 93.33 and 91.11% respectively for IFAT, ELISA and PCR.

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          Fluorescent antibody test for the serodiagnosis of American trypanosomiasis. Technical modification employing preserved culture forms of Trypanosoma cruzi in a slide test.

          M. E. Camargo (2015)
          Article 0 comments Cited 93 times – based on 0 reviews     Review now
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            Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (viannia) braziliensis.

            A de C Vexenat, J M Santana, A. R. Teixeira (2015)
            We have detected antibodies, in the sera of Chagas disease, Kala-azar and Mucocutaneous leishmaniasis patients, that bind multiple antigens shared between the three causative agents. The Chagas disease sera showed 98 to 100% positive results by ELISA when the Leishmania braziliensis and Leishmania chagasi antigens were used, respectively. The Kala-azar sera showed 100% positive results with Trypanosoma cruzi or L. braziliensis antigens by immunofluorescence assays. The antibodies in the sera of Mucocutaneous leishmaniasis patients showed 100% positive results by ELISA assays with T. cruzi or L. chagasi antigens. Furthermore, the direct agglutination of L. chagasi promastigotes showed that 95% of Kala-azar and 35% of Mucocutaneous leishmaniasis sera agglutinated the parasite in dilutions above 1:512. In contrast, 15% of Chagas sera agglutinated the parasite in dilutions 1:16 and below. Western blot analysis showed that the Chagas sera that formed at least 24 bands with the T. cruzi also formed 13 bands with the L. chagasi and 17 bands with the L. braziliensis. The Kala-azar sera that recognized at least 29 bands with the homologous antigen also formed 14 bands with the T. cruzi and 10 bands with the L. braziliensis antigens. Finally, the Mucocutaneous leishmaniasis sera that formed at least 17 bands with the homologous antigen also formed 10 bands with the T. cruzi and four bands with the L. chagasi antigens. These results indicate the presence of common antigenic determinants in several protozoal proteins and, therefore, explain the serologic cross-reactions reported here.
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              Leishmania spp. and/or Trypanosoma cruzi diagnosis in dogs from endemic and nonendemic areas for canine visceral leishmaniasis.

              Marcella Zampoli Troncarelli, Janaína Camargo, Juliana Machado … (2009)
              Due to the phylogenetic similarity between Leishmania spp. and Trypanosoma cruzi (T. cruzi), serological cross-reactions and false-positive results are quite common. This study aimed to elucidate canine leishmaniasis and trypanosomiasis diagnosis by the indirect fluorescent antibody test (IFAT) on serum samples, and direct parasitological examination and polymerase chain reaction (PCR) in liver and spleen samples. One hundred dogs from Zoonosis Control Center (ZCC) in Bauru, SP, an endemic area for visceral leishmaniasis (VL), and 100 dogs from the Dog Warden Service in Botucatu, SP, a nonendemic area for VL, were studied. IFAT showed positive results for Leishmania spp. in 65% of canine serum samples from Bauru while 40% of the samples were positive for T. cruzi by this test. All samples from Botucatu were negative for leishmaniasis in IFAT, and only 4% were positive for T. cruzi. Out of 200 serum samples tested, 33 (16.5%) showed positive serological results for both the parasites. Direct parasitological examination and PCR found, respectively, 59% and 76% of the liver samples and 51% and 72% of the spleen samples of dogs from Bauru positive for Leishmania spp. Twenty-six (78.8%) of 33 dogs that showed anti-Leishmania spp. and anti-T. cruzi antibodies also tested positive by direct parasitological examination and PCR for Leishmania spp., which indicates that these dogs presented leishmaniasis. No liver or spleen sample from the 200 dogs analyzed showed a positive PCR result for T. cruzi. These findings support the occurrence of cross-reactions between Leishmania spp. and T. cruzi in IFAT; they also corroborate the need for simultaneous PCR and/or parasitological examination to establish canine leishmaniasis (CL) diagnosis.
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                Author and article information

                Contributors
                JB Camargo: Role: ND
                H Langoni: Role: ND
                MZ Troncarelli: Role: ND
                JG Machado: Role: ND
                SB Lucheis: Role: ND
                CR Padovani: Role: ND
                Journal
                Journal ID (publisher): jvatitd
                Title: Journal of Venomous Animals and Toxins including Tropical Diseases
                Abbreviated Title: J. Venom. Anim. Toxins incl. Trop. Dis
                Publisher: Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESP (Botucatu )
                ISSN: 1678-9199
                Publication date (Print): 2010
                Volume: 16
                Issue: 3
                Pages: 414-420
                Affiliations
                [1 ] Universidade Estadual Paulista Brazil
                [2 ] Agência Paulista de Tecnologia dos Agronegócios Brazil
                [3 ] Universidade Estadual Paulista Brazil
                Article
                Publisher ID: S1678-91992010000300007
                DOI: 10.1590/S1678-91992010000300007
                SO-VID: 647d9c5e-eae6-4510-a5bd-636dd180b0d1
                License:

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1678-9199&lng=en
                Categories
                Subject: TOXICOLOGY
                Subject: TROPICAL MEDICINE

                ScienceOpen disciplines: Toxicology,Infectious disease & Microbiology
                Keywords: visceral leishmaniasis,dog,diagnosis
                Data availability:
                ScienceOpen disciplines: Toxicology, Infectious disease & Microbiology
                Keywords: visceral leishmaniasis, dog, diagnosis

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