Diagnóstico da leishmaniose visceral canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associação com a Rifi e Elisa-teste
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Data
2008-12-18
Autores
Queiroz, Nina Marí Gual Pimenta de [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A leishmaniose visceral canina (LVC) é causada pelo protozoário Leishmania (L.) chagasi e a presença deste parasita na pele dos cães reforça a importância destes animais no ciclo de vida da L. (L..) chagasi e na infecção humana. O objetivo deste trabalho foi avaliar o teste imunoistoquímico (IMIQ) e a PCR (Reação em Cadeia da Polimerase) em tecidos cutâneos caninos para o diagnóstico da LVC e compará-los com os exames histoquímico (HE) e sorológicos (ELISA e RIFI). Dos 34 cães naturalmente infectados, classificados em assintomáticos, oligossintomáticos e polissintomáticos, foram colhidas amostras de pele sadia ou com lesão para a realização da IMIQ, HE e PCR. Não somente peles lesionadas (56,5%), mas também peles sadias (31,8%) encontravam-se positivas pela IMIQ, confirmadas posteriormente pela PCR em 97,8% das amostras. No grupo assintomático, 87,5% estavam negativos pelos testes sorológicos, mas positivos em 50% dos casos pela IMIQ e 100% pela PCR. Entre os oligossintomáticos, 100%, 85,7% e 28,6% encontravam-se positivos, respectivamente pela PCR, sorologia e IMIQ. 91,7% dos polissintomáticos eram soro positivos e tinham parasitas intactos na pele. A PCR apresentou maior positividade, detectando amastigotas na pele de 100% dos cães independente do quadro clínico, do resultado dos exames sorológicos e do teste HE. A eficiência de cada teste variou de acordo com a evolução da doença, demonstrando a necessidade da associação de técnicas, usando IMIQ para confirmação da sorologia e a PCR apenas nos casos ainda suspeitos após a IMIQ, podendo assim aumentar os níveis de positividade e contribuir para o controle desta zoonose.
Canine visceral leishmaniasis (CVL) is caused by a protozoa of the specie Leishmania (L.) chagasi and the presence of parasite in the skin reinforces the importance of dogs in the L. (L.). chagasi life cycle and for human infection. The purpose of the present study was to evaluate the immunohistichemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for CVL diagnosis and compared the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitalogical test (microscopic direct examination of the parasite stained with haematoxylin and eosin - HE). For this study, samples of healthy or lesion skin tissues were obtained from 34 CVL naturally infected dogs classified in three groups: asymptomatic, oligosymptomatic and polisymptomatic. Not only lesion (56.5%) but also healthy skins (31.8%) were positives by IMHC and confirmed by PCR in 97.8% of skin samples. In asymptomatic group, 87.5% dogs were negatives by serological tests, but positives by IMHC in 50% and by PCR in 100%. In oligosymptomatic group, 100%, 85.7% and 28.6% of dogs were positives, respectively by PCR, serological and IMHC tests. In addition, 91.7% of polisymptomatic dogs were serum positives and had intact parasites in the skin. PCR showed higher positivity, detecting amastigotes in skins of 100% dogs regardless their clinical status or the serological or IMHC or HE test results. The efficiency of each test varied in agreement with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR only in suspicious cases after IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this zoonoses.
Canine visceral leishmaniasis (CVL) is caused by a protozoa of the specie Leishmania (L.) chagasi and the presence of parasite in the skin reinforces the importance of dogs in the L. (L.). chagasi life cycle and for human infection. The purpose of the present study was to evaluate the immunohistichemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for CVL diagnosis and compared the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitalogical test (microscopic direct examination of the parasite stained with haematoxylin and eosin - HE). For this study, samples of healthy or lesion skin tissues were obtained from 34 CVL naturally infected dogs classified in three groups: asymptomatic, oligosymptomatic and polisymptomatic. Not only lesion (56.5%) but also healthy skins (31.8%) were positives by IMHC and confirmed by PCR in 97.8% of skin samples. In asymptomatic group, 87.5% dogs were negatives by serological tests, but positives by IMHC in 50% and by PCR in 100%. In oligosymptomatic group, 100%, 85.7% and 28.6% of dogs were positives, respectively by PCR, serological and IMHC tests. In addition, 91.7% of polisymptomatic dogs were serum positives and had intact parasites in the skin. PCR showed higher positivity, detecting amastigotes in skins of 100% dogs regardless their clinical status or the serological or IMHC or HE test results. The efficiency of each test varied in agreement with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR only in suspicious cases after IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this zoonoses.
Descrição
Palavras-chave
Leishmania, Cão, Serologia, Leishmania, Dogs, Serology
Como citar
QUEIROZ, Nina Marí Gual Pimenta de. Diagnóstico da leishmaniose visceral canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associação com a Rifi e Elisa-teste. 2008. 125 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Odontologia de Araçatuba, 2008.