Diagnóstico da Leishmaniose Visceral Canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associaÇão com a RIFI e ELISA-teste

Abstract

The purpose of the present study was to evaluate the immunohistochemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for Canine Visceral Leishmaniasis (CVL) diagnosis and compare the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitological 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 positive and had intact parasites in the skin. In general, PCR showed higher positivity (100%). The efficiency of each test varied with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR in inconclusive cases after HE and IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this canine disease.

Molecular and serological detection of Leishmania spp. in captive wild animals from Ilha Solteira, SP, Brazil

“…Although this animal did not show skin lesions, there was an indication that parasites were present in healthy skin tissue and could potentially be transmitted to sandflies through feeding. Queiroz et al (2010) showed similar results in asymptomatic domestic dogs 100% PCR-positive from healthy skin samples. Abranches et al (1991) observed that 20% of dogs naturally infected by L. (L.) infantum had parasites in the healthy skin regardless of the presence of lesions.…”

“…using serological and molecular methods in captive wild animals from a zoo located in Ilha Solteira, São Paulo, Brazil, an area where VL is endemic and affects about 9% of the 3,798 domestic canid population (ASSIS et al, 2008;TASCA et al, 2009;QUEIROZ et al, 2010).…”

Correlation between chronic inflammation, immunostaining and parasite load in the genital system of female dogs naturally infected with Leishmania infantum

“…(MOREIRA et al, 2007). Immunohistochemistry (IHC) (TAFURI et al, 2004), polymerase chain reaction (PCR) and real-time PCR (qPCR) (QUEIROZ et al, 2010) are currently the most specific tests for the detection of L. infantum.…”

Patologias genitais associadas à leishmaniose visceral canina

“…Os segmentos da genitália externa apresentaram uma maior frequência de detecção por PCR (SILVA et al, 2008), uma vez que a vulva é coberta por tecido cutâneo ricamente suprido de glândulas apócrinas e sebáceas (DELLMANN & BROWN, 1982) e a pele está entre as amostras de escolha para extração de DNA e PCR nos casos de LVC (QUEIROZ et al, 2010). Dessa forma, a sensibilidade e especificidade atingem patamares próximos a 100% (FERRER, 1999;LUVIZOTTO, 2005).…”

“…Sendo assim, o diagnóstico laboratorial pode ser realizado através do exame parasitológico direto, testes imunológicos, como imunofluorescência indireta (RIFI) ou ensaio de imunoadsorção enzimático (ELISA), métodos imunohistoquímicos e, mais recentemente, através de métodos moleculares de amplificação do ácido nucleico, como a Reação da Cadeia em Polimerase (PCR) e PCR quantitativo (Real Time PCR) (MOREIRA et al, 2007;QUEIROZ et al, 2010;SOLCÁ et al, 2012).…”