Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina

Autores
Almazán, María Cristina; Krolewiecki, Alejandro Javier; Hoyos, Carlos Lorenzo; Cajal, Silvana Pamela; Copa, Noemí Griselda; Fleitas, Pedro Emanuel; Barroso, Paola Andrea; Marco, Jorge Diego; Nasser, Julio Rubén; Gil, José Fernando
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Cutaneous leishmaniasis is endemic in the northwestern province of Salta in Argentina. Leishmania DNA from Giemsa-stained slides of up to 12 years in storage of patients from a University based reference center in Oran, Salta was characterized through PCR-Restriction Fragment Length Polymorphism (RFLP). One hundred smears from individuals with suspicious lesions and epidemiologic exposure plus microbiologic confirmation through positive microscopy were analyzed. The samples had been originally taken 2, 3, 5, 6 and 12 years prior to performing PCR-RFLP (20 samples from each year) and were maintained at room temperature in a tropical environment. All the cases were classified in a semiquantitative scale for amastigote density, and Leishmanin skin test (LST) results were included. DNA extraction was done applying lysis buffer with proteinase K, and then DNA was amplified with ribosomal internal transcribed spacer 1(ITS1) primers. PCR products were digested with HaeIII enzyme. All PCR positive smears (74/100) belonged to Viannia subgenus. A statistically significant directly proportional relationship between semiquantitative microscopy and PCR results was detected (p < 0.001). All patients had LST positive results (induration ≥ 5 mm), and the smears of those with positive but smaller induration (LST < 19 mm) had a higher proportion of positive PCR results. This study determined that smear age did not affect PCR positivity, which allows retrospective analyzes and suggests smears might be useful for molecular complementary diagnosis. Since Leishmania (Viannia) braziliensis is the main circulating species in the study area, determining Viannia subgenus in all analyzed samples confirms previous findings. PCR positivity showed statistically significant differences according to semiquantitative microscopy, highlighting the importance of parasite burden in the diagnostic sensitivity of the method. Considering that smears of patients with smaller LST induration were more positive in PCR, a negative smear from patients with positive LST response, but < 19 mm, could actually represent a false negative smear result.
Fil: Almazán, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Krolewiecki, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Hoyos, Carlos Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Cajal, Silvana Pamela. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Copa, Noemí Griselda. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Fleitas, Pedro Emanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Barroso, Paola Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Marco, Jorge Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Nasser, Julio Rubén. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Gil, José Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
67th Annual Meeting American Society of Tropical Medicine & Hygiene
New Orleans
Estados Unidos
American Society of Tropical Medicine & Hygiene
Materia
LEISHMANIASIS
GIEMSA-STAINED SLIDES
ORAN
SPECIES IDENTIFICATION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/161430
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/161430
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, ArgentinaAlmazán, María CristinaKrolewiecki, Alejandro JavierHoyos, Carlos LorenzoCajal, Silvana PamelaCopa, Noemí GriseldaFleitas, Pedro EmanuelBarroso, Paola AndreaMarco, Jorge DiegoNasser, Julio RubénGil, José FernandoLEISHMANIASISGIEMSA-STAINED SLIDESORANSPECIES IDENTIFICATIONhttps://purl.org/becyt/ford/3.3https://purl.org/becyt/ford/3Cutaneous leishmaniasis is endemic in the northwestern province of Salta in Argentina. Leishmania DNA from Giemsa-stained slides of up to 12 years in storage of patients from a University based reference center in Oran, Salta was characterized through PCR-Restriction Fragment Length Polymorphism (RFLP). One hundred smears from individuals with suspicious lesions and epidemiologic exposure plus microbiologic confirmation through positive microscopy were analyzed. The samples had been originally taken 2, 3, 5, 6 and 12 years prior to performing PCR-RFLP (20 samples from each year) and were maintained at room temperature in a tropical environment. All the cases were classified in a semiquantitative scale for amastigote density, and Leishmanin skin test (LST) results were included. DNA extraction was done applying lysis buffer with proteinase K, and then DNA was amplified with ribosomal internal transcribed spacer 1(ITS1) primers. PCR products were digested with HaeIII enzyme. All PCR positive smears (74/100) belonged to Viannia subgenus. A statistically significant directly proportional relationship between semiquantitative microscopy and PCR results was detected (p < 0.001). All patients had LST positive results (induration ≥ 5 mm), and the smears of those with positive but smaller induration (LST < 19 mm) had a higher proportion of positive PCR results. This study determined that smear age did not affect PCR positivity, which allows retrospective analyzes and suggests smears might be useful for molecular complementary diagnosis. Since Leishmania (Viannia) braziliensis is the main circulating species in the study area, determining Viannia subgenus in all analyzed samples confirms previous findings. PCR positivity showed statistically significant differences according to semiquantitative microscopy, highlighting the importance of parasite burden in the diagnostic sensitivity of the method. Considering that smears of patients with smaller LST induration were more positive in PCR, a negative smear from patients with positive LST response, but < 19 mm, could actually represent a false negative smear result.Fil: Almazán, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Krolewiecki, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Hoyos, Carlos Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Cajal, Silvana Pamela. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Copa, Noemí Griselda. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Fleitas, Pedro Emanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Barroso, Paola Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Marco, Jorge Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Nasser, Julio Rubén. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; ArgentinaFil: Gil, José Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina67th Annual Meeting American Society of Tropical Medicine & HygieneNew OrleansEstados UnidosAmerican Society of Tropical Medicine & HygieneAmerican Society of Tropical Medicine & Hygiene2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/161430Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina; 67th Annual Meeting American Society of Tropical Medicine & Hygiene; New Orleans; Estados Unidos; 2018; 441-441CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.astmh.org/annual-meeting/past-meetingsInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:07:08Zoai:ri.conicet.gov.ar:11336/161430instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:07:09.01CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
title Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
spellingShingle Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
Almazán, María Cristina
LEISHMANIASIS
GIEMSA-STAINED SLIDES
ORAN
SPECIES IDENTIFICATION
title_short Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
title_full Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
title_fullStr Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
title_full_unstemmed Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
title_sort Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina
dc.creator.none.fl_str_mv Almazán, María Cristina
Krolewiecki, Alejandro Javier
Hoyos, Carlos Lorenzo
Cajal, Silvana Pamela
Copa, Noemí Griselda
Fleitas, Pedro Emanuel
Barroso, Paola Andrea
Marco, Jorge Diego
Nasser, Julio Rubén
Gil, José Fernando
author Almazán, María Cristina
author_facet Almazán, María Cristina
Krolewiecki, Alejandro Javier
Hoyos, Carlos Lorenzo
Cajal, Silvana Pamela
Copa, Noemí Griselda
Fleitas, Pedro Emanuel
Barroso, Paola Andrea
Marco, Jorge Diego
Nasser, Julio Rubén
Gil, José Fernando
author_role author
author2 Krolewiecki, Alejandro Javier
Hoyos, Carlos Lorenzo
Cajal, Silvana Pamela
Copa, Noemí Griselda
Fleitas, Pedro Emanuel
Barroso, Paola Andrea
Marco, Jorge Diego
Nasser, Julio Rubén
Gil, José Fernando
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv LEISHMANIASIS
GIEMSA-STAINED SLIDES
ORAN
SPECIES IDENTIFICATION
topic LEISHMANIASIS
GIEMSA-STAINED SLIDES
ORAN
SPECIES IDENTIFICATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.3
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Cutaneous leishmaniasis is endemic in the northwestern province of Salta in Argentina. Leishmania DNA from Giemsa-stained slides of up to 12 years in storage of patients from a University based reference center in Oran, Salta was characterized through PCR-Restriction Fragment Length Polymorphism (RFLP). One hundred smears from individuals with suspicious lesions and epidemiologic exposure plus microbiologic confirmation through positive microscopy were analyzed. The samples had been originally taken 2, 3, 5, 6 and 12 years prior to performing PCR-RFLP (20 samples from each year) and were maintained at room temperature in a tropical environment. All the cases were classified in a semiquantitative scale for amastigote density, and Leishmanin skin test (LST) results were included. DNA extraction was done applying lysis buffer with proteinase K, and then DNA was amplified with ribosomal internal transcribed spacer 1(ITS1) primers. PCR products were digested with HaeIII enzyme. All PCR positive smears (74/100) belonged to Viannia subgenus. A statistically significant directly proportional relationship between semiquantitative microscopy and PCR results was detected (p < 0.001). All patients had LST positive results (induration ≥ 5 mm), and the smears of those with positive but smaller induration (LST < 19 mm) had a higher proportion of positive PCR results. This study determined that smear age did not affect PCR positivity, which allows retrospective analyzes and suggests smears might be useful for molecular complementary diagnosis. Since Leishmania (Viannia) braziliensis is the main circulating species in the study area, determining Viannia subgenus in all analyzed samples confirms previous findings. PCR positivity showed statistically significant differences according to semiquantitative microscopy, highlighting the importance of parasite burden in the diagnostic sensitivity of the method. Considering that smears of patients with smaller LST induration were more positive in PCR, a negative smear from patients with positive LST response, but < 19 mm, could actually represent a false negative smear result.
Fil: Almazán, María Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Krolewiecki, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Hoyos, Carlos Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Cajal, Silvana Pamela. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Copa, Noemí Griselda. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Fleitas, Pedro Emanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Barroso, Paola Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Marco, Jorge Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Nasser, Julio Rubén. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
Fil: Gil, José Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Universidad Nacional de Salta. Sede Regional Orán. Instituto de Investigación de Enfermedades Tropicales; Argentina
67th Annual Meeting American Society of Tropical Medicine & Hygiene
New Orleans
Estados Unidos
American Society of Tropical Medicine & Hygiene
description Cutaneous leishmaniasis is endemic in the northwestern province of Salta in Argentina. Leishmania DNA from Giemsa-stained slides of up to 12 years in storage of patients from a University based reference center in Oran, Salta was characterized through PCR-Restriction Fragment Length Polymorphism (RFLP). One hundred smears from individuals with suspicious lesions and epidemiologic exposure plus microbiologic confirmation through positive microscopy were analyzed. The samples had been originally taken 2, 3, 5, 6 and 12 years prior to performing PCR-RFLP (20 samples from each year) and were maintained at room temperature in a tropical environment. All the cases were classified in a semiquantitative scale for amastigote density, and Leishmanin skin test (LST) results were included. DNA extraction was done applying lysis buffer with proteinase K, and then DNA was amplified with ribosomal internal transcribed spacer 1(ITS1) primers. PCR products were digested with HaeIII enzyme. All PCR positive smears (74/100) belonged to Viannia subgenus. A statistically significant directly proportional relationship between semiquantitative microscopy and PCR results was detected (p < 0.001). All patients had LST positive results (induration ≥ 5 mm), and the smears of those with positive but smaller induration (LST < 19 mm) had a higher proportion of positive PCR results. This study determined that smear age did not affect PCR positivity, which allows retrospective analyzes and suggests smears might be useful for molecular complementary diagnosis. Since Leishmania (Viannia) braziliensis is the main circulating species in the study area, determining Viannia subgenus in all analyzed samples confirms previous findings. PCR positivity showed statistically significant differences according to semiquantitative microscopy, highlighting the importance of parasite burden in the diagnostic sensitivity of the method. Considering that smears of patients with smaller LST induration were more positive in PCR, a negative smear from patients with positive LST response, but < 19 mm, could actually represent a false negative smear result.
publishDate 2019
dc.date.none.fl_str_mv 2019
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/161430
Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina; 67th Annual Meeting American Society of Tropical Medicine & Hygiene; New Orleans; Estados Unidos; 2018; 441-441
CONICET Digital
CONICET
url http://hdl.handle.net/11336/161430
identifier_str_mv Molecular characterization of Leishmania DNA from archived giemsa-stained slides of patients from Salta, Argentina; 67th Annual Meeting American Society of Tropical Medicine & Hygiene; New Orleans; Estados Unidos; 2018; 441-441
CONICET Digital
CONICET
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language eng
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rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv American Society of Tropical Medicine & Hygiene
publisher.none.fl_str_mv American Society of Tropical Medicine & Hygiene
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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