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

Autor:	Garay AFG; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Fraenkel S; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Diaz JJAR; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Recalde ODS; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Gómez MCV; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Riquelme JAM; Ministerio de Salud Pública y Bienestar Social, Programa Nacional de Control de Zoonosis y Centro Antirrábico Nacional, San Lorenzo, Central, Paraguay.; Universidad Nacional de Asunción, Instituto de Investigaciones en Ciencias de la Salud, Departamento de Medicina Tropical, San Lorenzo, Central, Paraguay., Arze PV; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Centurión GNR; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Britos M; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay., Rolón M; Centro para el Desarrollo de la Investigación Científica, Asunción, Central, Paraguay.
Jazyk:	angličtina
Zdroj:	Revista da Sociedade Brasileira de Medicina Tropical [Rev Soc Bras Med Trop] 2022 Dec 16; Vol. 55. Date of Electronic Publication: 2022 Dec 16 (Print Publication: 2022).
DOI:	10.1590/0037-8682-0069-2022
Abstrakt:	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.
Databáze:	MEDLINE
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