Diagnostic Techniques To Detect Cryptic Leishmaniasis in Dogs

Iniesta, Laura; Fernández-Barredo, Salceda; Bulle, Béatrice; Gómez, Manel; Piarroux, Renaud; Gállego, Montserrat; Alunda, José María; Portús, Montserrat

doi:10.1128/cdli.9.5.1137-1141.2002

Cited by 40 publications

(51 citation statements)

References 26 publications

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“…The low sensitivity of in vitro culture observed in our study is in correlation with what is found in some studies (Berrahal et al, 1996, Fisa et al, 2001. However, other studies found sensitivity as high as that of PCR in dog blood (Iniesta et al, 2002) or in human blood (Kaouach et al, 2008). This difference can be due to the method used and the local conditions of in vitro culture.…”

Section: Diagnosis Of Canine Leishmaniasismentioning

confidence: 93%

Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

et al. 2009

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Summary :The aim of this study was to assess the use of parasitological, serological and molecular methods for the detection of Leishmania infection in blood of 67 dogs and to investigate the prevalence of canine leishmaniasis (CanL) in Kairouan (central Tunisia), an area known to be of reduced endemicity and has not been studied since 1973. Veterinarians clinically examined all dogs, and the titer of anti-Leishmania antibodies was determined by indirect immune-fluorescence antibody test. The presence of Leishmania was performed by PCR and in vitro culture. IFAT was positive in 12 % of dogs and promastigote form of the parasite was isolated by in vitro culture from only 4.5 % of them. However, DNA of Leishmania was detected by PCR in 20.9 % of dogs. PCR was more sensitive than IFAT (p = 0.004) and in vitro culture (p < 10 -5 ). A prevalence of 21 % was found in Kairouan, which is significant high (p < 10 -3 ) when compared to that of thirty years ago. This state is in correlation with the increase in other Mediterranean countries. Furthermore, 50 % of positive dogs were asymptomatic. Preventive measures must be taken against these dogs as for symptomatic ones since their role in the transmission of the infection to vectors has been proven.

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Section: Diagnosis Of Canine Leishmaniasismentioning

confidence: 93%

Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

et al. 2009

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“…The presence of L. infantum kDNA in the blood of most of our patients did not lead to either a cellular or a humoral response in our patients. Surveys conducted to detect cryptic leishmaniasis in dogs also showed a higher prevalence by PCRpositive results than by antibody detection techniques (9,22). Le Fichoux et al (10) suggested that in healthy L. infantum carriers, episodes of parasitemia might be of low intensity and, therefore, a cellular or humoral immune response may not be triggered.…”

Section: Discussionmentioning

confidence: 99%

Use of Noninvasive Markers To Detect Leishmania Infection in Asymptomatic Human Immunodeficiency Virus-Infected Patients

et al. 2006

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Visceral leishmaniasis (VL) caused by Leishmania infantum is a common disease in human immunodeficiency virus (HIV)-infected people in the Mediterranean basin. However, most such cases are asymptomatic, and little information about the prevalence of these infections in HIV-infected individuals is available. The aim of this study was to assess the prevalence of subclinical infection and the relationship between several Leishmania infection markers by noninvasive methods in asymptomatic HIV-infected patients from Southern Spain. Ninety-two HIV-infected patients, who were consecutively attended at the participant hospitals in 2004, were invited to participate in this study. These patients were asymptomatic and without any history of cutaneous or visceral leishmaniasis. Leishmania kinetoplast DNA (kDNA) was amplified from peripheral blood samples from 28 (30.4%) of these HIV-infected subjects. Sera from three (3.5%) patients tested positive for Leishmania by an enzyme-linked immunoassay. Two patients (2.4%) showed a specific 16-kDa band by Western blotting. In contrast, none of the patients showed a positive agglutination of urine. The leishmanin skin test was positive for four (4.3%) patients. None of the patients with a PCR-positive result showed a positive reaction by enzyme-linked immunoassay or by specific bands in Western blotting or had a positive leishmanin skin test. In conclusion, L. infantum kDNA was detected in a large proportion of asymptomatic HIV-infected patients, although a demonstrable cellular or humoral immune response to this parasite was not shown. Conversely, Leishmania antigen in urine was not detected in these patients.Visceral leishmaniasis (VL) caused by Leishmania infantum is a common coinfection in human immunodeficiency virus (HIV)-infected people in Spain (3), although most patients remain asymptomatic (15). Tissue culture or direct examination shows Leishmania promastigotes or amastigotes, respectively, in a considerable proportion of asymptomatic HIVinfected subjects (15). However, invasive techniques that are not often well tolerated by asymptomatic individuals are required to obtain these samples.Recently, a number of noninvasive methods that are easier to perform and better tolerated have been developed for the diagnosis of leishmaniasis and asymptomatic VL. Serology and leishmanin skin tests (LST) are easy to use but have low sensitivity in HIV-and Leishmania-coinfected patients (3, 14). More recently, PCR-based methods for detecting Leishmania species have been used for testing peripheral blood samples (7,8,13,21). In addition, diagnostic techniques have also been developed for the identification of Leishmania antigen in urine (4,6,23,26,27). Some studies have been undertaken in order to determine the prevalence of Leishmania infection in blood donors or asymptomatic individuals (5,11,21). However, the prevalence of asymptomatic VL in HIV-infected subjects is not well known. Moreover, little is known about the relationships between the results obtained by both PCR or urine antig...

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“…Para o diagnóstico da LVC, as técnicas sorológicas são consideradas rápidas e práticas, mas com sensibilidade variável e especificidade baixa (BADARÓ et al, 1983;INIESTA et al, Tabela 3. Análise comparativa do número (nc) e porcentagens (%) de concordância e índice Kappa (κ) entre as técnicas imunoistoquímica (IMIQ) × ELISA, IMIQ × RIFI, ELISA × RIFI, PCR × IMIQ, PCR × ELSA e PCR × RIFI para diagnóstico de Leishmaniose Visceral em cães naturalmente infectados e classificados em assintomáticos (A), oligossintomáticos (O) e polissintomáticos (P).…”

Section: Grupos (N)unclassified

“…Enquanto os testes sorológicos apresentam mais resultados falso positivos por causa das reações cruzadas com outros patógenos (BADARÓ et al, 1983;INIESTA et al, 2002), os métodos parasitológicos são mais precisos, mas também mais invasivos porque, usualmente, requerem punção de linfonodo periférico ou medula óssea.…”

Section: Introductionunclassified

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

Queiroz¹,

Assis²,

Oliveira³

et al. 2010

Rev. Bras. Parasitol. Vet. (Online)

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ResumoO objetivo deste trabalho foi avaliar as técnicas de imunoistoquímica (IMIQ) e de PCR (Reação em Cadeia da Polimerase) em tecidos cutâneos para o diagnóstico da Leishmaniose Visceral Canina (LVC) e compará-los com os exames parasitológicos em tecidos corados histoquimicamente (hematoxilina-eosina, HE) e com testes sorológicos, como a Reação de Imunofluorescência Indireta (RIFI) e ensaio imunoenzimático (ELISA). Dos 34 cães naturalmente infectados, classificados em assintomáticos, oligossintomáticos e polissintomáticos, foram coletadas amostras de pele sadia ou com lesão para a realização da IMIQ, HE e PCR. Não somente peles lesionadas (56,5%), mas também 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. Os cães polissintomáticos eram 91,7% soropositivos e tinham parasitas na pele. Em geral, a técnica PCR teve maior positividade (100%). A eficiência dos testes variou de acordo com a evolução da doença, demonstrando a necessidade da associação de técnicas, usando-se IMIQ para confirmação da sorologia e a PCR apenas nos casos suspeitos após a IMIQ. Dessa forma, pode-se aumentar os níveis de positividade e contribuir para o controle desta zoonose.Palavras-chave: Leishmania, cães, pele, imunoistoquímica, PCR. AbstractThe 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.

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Diagnostic Techniques To Detect Cryptic Leishmaniasis in Dogs

Cited by 40 publications

References 26 publications

Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

Use of PCR, IFAT andin vitroculture in the detection ofLeishmania infantuminfection in dogs and evaluation of the prevalence of canine leishmaniasis in a low endemic area in Tunisia

Use of Noninvasive Markers To Detect Leishmania Infection in Asymptomatic Human Immunodeficiency Virus-Infected Patients

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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