Estudo de validação comparativo entre as técnicas Elisa e RIFI para diagnosticar Leishmania sp em cães errantes apreendidos no município de Campos dos Goytacazes, Estado do Rio de Janeiro

Távora, Mariane Pinto Fernandes; Pereira, Maria Angélica Vieira da Costa; Silva, Valmir Laurentino; Vita, Gilmar Ferreira

doi:10.1590/s0037-86822007000400023

Cited by 6 publications

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“…Both of these tests were available through the freeware BioEstat version 5.0, and were used with 5% statistical significance. Co-positivity was calculated as a/(a + c), co-negativity was expressed as d/(b + b) and raw concordance was described as (a + d)/(a + b + c + d) (TÁVORA et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

Molecular and parasitological detection of Leishmania spp. in dogs caught in Palmas, TO, Brazil

Teles

Agostini

Bigeli

et al. 2012

Rev. Bras. Parasitol. Vet.

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This study evaluated occurrences of Leishmania infantum in dogs in the municipality of Palmas, Tocantins, comparing diagnostic data obtained using the polymerase chain reaction (PCR) and parasitological diagnosis. Blood samples and lymph node aspirates were collected from 63 dogs of males and females and various ages and races, with or without owners, between August 2009 and June 2010. Slides containing smears of lymph node aspirates were stained with Giemsa stained. In PCR, the 145 bp target sequence of the LT1 fragment, located in the Leishmania donovani kDNA minicircle was detected using the RV1 and RV2 oligonucleotide primers. The chi-square test revealed that there was a significant relationship between the symptoms and dogs that were positive for visceral leishmaniasis (VL). The parasitological investigation showed concordance of 66.7% with PCR on blood and 84.1% with PCR on lymph node aspirate. In addition to these tests, evaluations of the diagnoses in parallel and in series were conducted, which showed concordances with the parasitological test of 76.2% and 74.6%, respectively. The results make it possible to suggest that PCR on lymph nodes should be used in evaluating large populations (surveys) and that the parasitological test should be used for initial clinical evaluations in veterinary consultation offices.Keywords: PCR, blood, lymph node aspirate, parasite, diagnosis. ResumoAvaliou-se a ocorrência de Leishmania infantum em cães do município de Palmas-TO, comparando dados diagnósticos obtidos pela Reação em Cadeia da Polimerase (PCR) e pelo diagnóstico parasitológico. Foram coletadas amostras de sangue e de aspirado de linfonodo de 63 cães machos e fêmeas, várias idades e raças, domiciliares ou não de agosto de 2009 a junho de 2010. As lâminas contendo esfregaço dos aspirados de linfonodos foram coradas pelo corante Giemsa. Na PCR, a sequência alvo de 145 pb do fragmento LT1, situado no minicírculo do kDNA do grupo Leishmania donovani, foi detectada através dos oligonucleotídeos iniciadores RV1 e RV2. O teste χ 2 (Qui-quadrado), demonstrou haver relação significativa entre a sintomatologia e a positividade dos cães para Leishmaniose Visceral (LV). O exame parasitológico mostrou uma concordância de 66,7% com a PCR em sangue e 84,1% com a PCR de aspirado de linfonodo. Além destas análises, houve a avaliação dos diagnósticos em paralelo e em série, onde as concordâncias com o exame parasitológico foram de 76,2% e 74,6%, respectivamente. Os resultados permitem sugerir a utilização da PCR de linfonodos na avaliação de grandes populações (inquéritos), e o exame parasitológico para a avaliação clínica inicial em consultórios veterinários.Palavras-chaves: PCR, sangue, aspirado de linfonodos, parasita, diagnóstico.

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Section: Discussionmentioning

confidence: 99%

Molecular and parasitological detection of Leishmania spp. in dogs caught in Palmas, TO, Brazil

Teles

Agostini

Bigeli

et al. 2012

Rev. Bras. Parasitol. Vet.

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“…Low agreement between the serological tests in our study can also be observed in researches carried out in Monte Negro -Rondônia (RO) State (AGUIAR et al, 2010), between ELISA and IFAT test, in Panorama -São Paulo (SP) State (LOPES et al, 2017), between ELISA and DPP tests. However, also high and moderate agreements have already been observed between the ELISA and IFAT tests in other localities of Brazil, as in Campo do Goycatazes -Rio de Janeiro State (TÁVORA et al, 2007) and Ilha Solteira (SP) (ASSIS et al, 2010;QUEIROZ et al, 2010). These observations demonstrated the need to better assess the diagnostic techniques of CVL, aiming at standardization, considering the evolution of the disease in the dog and its immune response.…”

Section: Discussionmentioning

confidence: 86%

Canine visceral leishmaniosis in an area of fishing tourism, Bonito Municipality, Mato Grosso do Sul, Central-West Brazil

et al. 2018

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The study aimed to investigate Leishmania infection in a population of dogs (92 animals) in a fishing area of Bonito Municipality, and evaluate comparatively the serological methods used, immunoenzymatic assay (ELISA), indirect immunofluorescence antibody test (IFAT), and rapid Dual Path-Platform test (DPP®). Blood and aspirate of bone marrow samples were used and parasitological investigation was also performed, such as parasite isolation in Neal, Novy, Nicolle (NNN) medium culture, Woo technique, Giemsa stained smears and specific identification by polymerase chain reaction (PCR). IFAT revealed 56/92 seropositive, the ELISA 8/92 and the DPP®41/92. Regarding the agreement between the serological tests by the Kappa index, there was a slight agreement between ELISA/IFAT and ELISA/DPP®, and fair agreement in IFAT/DPP®. The NNN culture was positive in three out of 21 dogs, and identified by PCR as Leishmania infantum chagasi. No samples were positive by the Woo technique. Our results showed low agreements between the serological tests recommended by the Brazilian Ministry of Heath, and it is necessary to associate other diagnostic techniques, such as parasitological tests and PCR, to increase the sensitivity for canine visceral leishmaniosis diagnosis, mainly regarding asymptomatic dogs in endemic areas.

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“…In previous studies, cross-reactivity between Leishmania species and T. gondii was tested by ELISA and IFAT (Boelaert et al, 2007;Ferreira et al, 2007) and showed no cross-reactivity. However, other studies showed that half of the dogs (5/10; 50%) with anti-T. gondii antibodies were erroneously considered serologically positive for VL (Távora et al, 2007). In evaluating the validity of diagnostic methods, such problems need to be taken fully into account in order to obtain reliable and accurate results.…”

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confidence: 99%

The efficacy of recombinant protein lbk39 for the diagnosis of leishmaniosis in dogs

et al. 2020

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Visceral leishmaniosis is one of the most important zoonotic diseases on the planet and dogs are the main reservoir of canine visceral leishmaniosis (CVL) in endemic areas. They play an important role in human infection because in dogs the disease appears long time after infection, and they can move uncontrollably, contributing to disperse the parasite. To take the decision to treat the animals or for euthanasia, in an elimination programme, in order to reduce the parasitic load, it is necessary to diagnose correctly, having more effective tools. Our group has developed a new recombinant antigen-based kinesin-related gene of Leishmania braziliensis (Lbk39), which shows 59% amino acid identity to the L. infantum homologue. The Lbk39 gene was synthesized, inserted into the pLEXSY-sat2 vector and transfected into L. tarentolae cells by electroporation. The recombinant protein was secreted in the culture with a C-terminal histidine marker, purified, generating a product at 337.68 μg mL−1. A total of 152 sera from dog's endemic and non-endemic areas were used, being 78 positives and 75 negatives. The antigen Lbk39 showed 100% sensitivity and 96.1% specificity. We compared this antigen with other antigens such as total extract of the parasite, TRDPP, and our data indicate that Lbk39 has potential application in the diagnosis of CVL through antibody detection.

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Estudo de validação comparativo entre as técnicas Elisa e RIFI para diagnosticar Leishmania sp em cães errantes apreendidos no município de Campos dos Goytacazes, Estado do Rio de Janeiro

Cited by 6 publications

References 0 publications

Molecular and parasitological detection of Leishmania spp. in dogs caught in Palmas, TO, Brazil

Molecular and parasitological detection of Leishmania spp. in dogs caught in Palmas, TO, Brazil

Canine visceral leishmaniosis in an area of fishing tourism, Bonito Municipality, Mato Grosso do Sul, Central-West Brazil

The efficacy of recombinant protein lbk39 for the diagnosis of leishmaniosis in dogs

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