Comparative PCR-based diagnosis for the detection of Leishmania infantum in naturally infected dogs.

Marcelino, Andreza Pain; Filho, Job Alves de Souza; Bastos, Camila de Valgas e; Ribeiro, Simone Rodrigues; Medeiros, Fernanda Alvarenga Cardoso; Reis, Ilka Afonso; Lima, Ana Cristina Vianna Mariano da Rocha; Barbosa, José Ronaldo; Paz, Gustavo Fontes; Gontijo, Célia Maria Ferreira

doi:10.1016/j.actatropica.2020.105495

Cited by 13 publications

(11 citation statements)

References 35 publications

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“…In the context of this study, the obtained results showed that the samples extracted from distinct types of nodes showed higher levels of sensitivity, as previously described 32 , 33 , in comparison to the BM and EP samples. Although the literature mentions that by using the same technique, the bone marrow samples present similar sensitivity values compared to the lymph nodes, the lower sensitivity values observed in different samples in this study could have been due to the low parasitic load in the tissues or associated issues with the obtainment or processing of the samples 34 .…”

Section: Discussionmentioning

confidence: 63%

Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM

Garay¹,

Fraenkel²,

Diaz³

et al. 2022

Rev. Soc. Bras. Med. Trop.

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Background: Leishmaniasis is a vector-borne disease caused by a parasite protozoon from the genus Leishmania. Among the molecular techniques applied for detecting these parasites, real-time PCR with High Resolution Melting (PCR-HRM) proved advantageous since it simultaneously determines both the presence and species of the pathogen in one step, through amplification and later analysis of curves generated by melting temperature. Methods:Based on this molecular technique, the goal of this study was to estimate the PCR-HRM sensitivity for Leishmania spp. detection in different canine tissues by evaluating biological samples obtained from popliteal, submandibular, and pre-scapular lymph nodes, from bone marrow and ear pinnae of 28 stray dogs captured in the metropolitan area of Asunción (Paraguay). Results:The rk39 immunochromatographic test showed that 25/28 tested dogs (89%) presented antibodies against L. infantum. In 20/25 dogs that tested positive for rk39 (80%), it was possible to detect Leishmania spp. by PCR-HRM and determine that the species corresponded entirely to L. infantum. Regarding the analysis of different tissues, the parasite was detected in all popliteal lymph node samples, followed by high detection in submandibular (at 95%) and pre-scapular lymph nodes (at 90%), bone marrow (at 85%), and ear pinnae (at 85%). Conclusions:This study demonstrated that the use of real-time PCR-HRM using the molecular marker hsp70 was a highly sensitive method for simultaneously detecting and identifying Leishmania species in different tissues taken from infected dogs. In addition, the usefulness of ear pinnae as easily accessible tissue for molecular diagnosis was emphasized.

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Section: Discussionmentioning

confidence: 63%

Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM

Garay¹,

Fraenkel²,

Diaz³

et al. 2022

Rev. Soc. Bras. Med. Trop.

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“…Considering the PCR results, 7.1% of the dogs were positive for the presence of L. infantum in the peripheral blood, although detection of L. infantum by PCR from blood samples (buffy coat) is less sensitive for CVL diagnosis than sampling bone marrow or popliteal lymph nodes (Silveira et al, 2018;Marcelino et al, 2020). Studies carried out in the states of Minas Gerais and Mato Grosso do Sul using blood samples revealed 24.7% and 47.5% of positive PCR, respectively (Coura-Vital et al, 2011;Cunha et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

First case of canine visceral leishmaniasis in the midwestern of Santa Catarina State, Brazil

et al. 2022

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Canine visceral leishmaniasis (CVL) caused by Leishmania (Leishmania) infantum is transmitted by phlebotomine sandflies and a major zoonotic disease in Brazil. Due to the southward expansion of the disease within the country and the central role of dogs as urban reservoirs of the parasite, we have investigated the occurrence of CVL in two municipalities Erval Velho and Herval d’Oeste in the Midwest region of Santa Catarina state. Peripheral blood samples from 126 dogs were collected in both cities and tested for anti-L. infantum antibodies by indirect enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence reaction (IIF) and for the presence of parasite DNA by polymerase chain reaction (PCR) in peripheral blood. From examined dogs, 35.71% (45/126) were positive for at least one of the three tests and two (1.6%) were positive in all performed tests. Twelve dogs (9.5%) were positive for both ELISA and IIF, while 21 dogs were exclusively positive for ELISA (16.7%), and 15 (11.9%) for IIF. L. infantum k-DNA was detected by PCR in 9 out of 126 dogs (7.1%) and clinical symptoms compatible with CVL were observed for 6 dogs. Taken together, these results indicate the transmission of CVL in this region, highlighting the needs for epidemiological surveillance and implementation of control measures for CVL transmission in this region.

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“…Sin embargo, a pesar de ser considerada una de las mejores opciones de diagnóstico en investigación, este tipo de técnicas presentan algunas desventajas que evitan su uso universal y estandarizado, como son: 1) la elección del órgano/fluido blanco para la muestra, ya que dependiendo de la etapa de la enfermedad (aguda o crónica en EC), la sensibilidad cambia, 2) la purificación del material para amplificar (DNA-RNA) se puede contaminar y generar resultados falsos, y 3) la elección adecuada de los oligonucleótidos para la reacción, puesto que existe una amplia variedad de genes que se pueden amplificar para identificar una especie. Por tanto, es necesario elegir aquel oligonucleótido que permita obtener resultados de alta especificidad y sensibilidad para la detección de microorganismos de interés (Kirchhoff et al, 1996;López-Céspedes, 2013;Marcelino et al, 2020). ¿Estándar de oro?…”

Section: Discussionunclassified

Actualización del estado de conocimiento sobre las técnicas de diagnóstico de la leishmaniasis y enfermedad de Chagas

Pineda-Rios,

González-Rete,

Cañeda-Guzmán

et al. 2024

Rev.Mex.Biodiv.

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La leishmaniasis y la enfermedad de Chagas son causadas por protozoarios que comparten características filogenéticas y eco-epidemiológicas comunes, lo cual provoca dificultad en su identificación y diagnóstico. El objetivo de este trabajo fue analizar la información disponible sobre las técnicas de identificación de T. cruzi, de Leishmania spp. y la coinfección con ambos parásitos, así como su impacto en la obtención de datos eco-epidemiológicos. Se realizó una búsqueda sistemática de técnicas de identificación para T. cruzi, Leishmania spp. o ambos. Los estudios se clasificaron por agente etiológico, tipo de publicación, técnica utilizada y hospedero. Se realizó una prueba de X2 para evaluar la relación entre el parásito y la técnica utilizada. También se analizó la relación entre los trabajos publicados por año, la técnica diagnóstica y el parásito. De 138 trabajos analizados, se determinó que no existe un “estándar de oro” para el diagnóstico de estas parasitosis y la coinfección. Por tanto, se requiere la estandarización de protocolos que incrementen la sensibilidad y especificidad, así como el uso de al menos 2 técnicas en la identificación. Es imprescindible estudiar la prevalencia e incidencia de estas parasitosis en la fauna silvestre y la población humana bajo el contexto de cambio climático.

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Comparative PCR-based diagnosis for the detection of Leishmania infantum in naturally infected dogs.

Cited by 13 publications

References 35 publications

Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM

Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM

First case of canine visceral leishmaniasis in the midwestern of Santa Catarina State, Brazil

Actualización del estado de conocimiento sobre las técnicas de diagnóstico de la leishmaniasis y enfermedad de Chagas

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