Cargando…
Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs
Canine leishmaniasis (CanL) diagnosis is not fully resolved. Currently, two specific methodologies are in continuous development, the detection of the parasite DNA or RNA in target organs and the detection of specific antibodies against Leishmania sp. For a correct diagnosis, it has been shown that...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8704913/ https://www.ncbi.nlm.nih.gov/pubmed/34946227 http://dx.doi.org/10.3390/microorganisms9122627 |
_version_ | 1784621820294463488 |
---|---|
author | Peris, María Paz Esteban-Gil, Adriana Ortega-Hernández, Paula Morales, Mariano Halaihel, Nabil Castillo, Juan Antonio |
author_facet | Peris, María Paz Esteban-Gil, Adriana Ortega-Hernández, Paula Morales, Mariano Halaihel, Nabil Castillo, Juan Antonio |
author_sort | Peris, María Paz |
collection | PubMed |
description | Canine leishmaniasis (CanL) diagnosis is not fully resolved. Currently, two specific methodologies are in continuous development, the detection of the parasite DNA or RNA in target organs and the detection of specific antibodies against Leishmania sp. For a correct diagnosis, it has been shown that the joint use of this type of test is necessary. In this work, a Sybr Green and a TaqMan Probe based on real time PCRs (qPCR) was performed for the detection of Leishmania sp. in order to correlate the results with clinicopathological and serological evaluations (IFA, ELISA and DAT) to propose an optimal biological sample to be used to detect the parasite in both early and late stages of the infection. A total of four samples were processed: conjunctival swabs, popliteal lymph node aspirates, bone marrow aspirates, and peripheral blood from experimentally infected dogs belonging to a larger study. Our results indicated that a single non-invasive sample (conjunctival swab) and the application of both types of qPCR would be reliable for determining Leishmania infection as well as the disease stage in dogs, thus avoiding bone marrow, lymph node aspirate or blood samples collection. |
format | Online Article Text |
id | pubmed-8704913 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-87049132021-12-25 Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs Peris, María Paz Esteban-Gil, Adriana Ortega-Hernández, Paula Morales, Mariano Halaihel, Nabil Castillo, Juan Antonio Microorganisms Article Canine leishmaniasis (CanL) diagnosis is not fully resolved. Currently, two specific methodologies are in continuous development, the detection of the parasite DNA or RNA in target organs and the detection of specific antibodies against Leishmania sp. For a correct diagnosis, it has been shown that the joint use of this type of test is necessary. In this work, a Sybr Green and a TaqMan Probe based on real time PCRs (qPCR) was performed for the detection of Leishmania sp. in order to correlate the results with clinicopathological and serological evaluations (IFA, ELISA and DAT) to propose an optimal biological sample to be used to detect the parasite in both early and late stages of the infection. A total of four samples were processed: conjunctival swabs, popliteal lymph node aspirates, bone marrow aspirates, and peripheral blood from experimentally infected dogs belonging to a larger study. Our results indicated that a single non-invasive sample (conjunctival swab) and the application of both types of qPCR would be reliable for determining Leishmania infection as well as the disease stage in dogs, thus avoiding bone marrow, lymph node aspirate or blood samples collection. MDPI 2021-12-20 /pmc/articles/PMC8704913/ /pubmed/34946227 http://dx.doi.org/10.3390/microorganisms9122627 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Peris, María Paz Esteban-Gil, Adriana Ortega-Hernández, Paula Morales, Mariano Halaihel, Nabil Castillo, Juan Antonio Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs |
title | Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs |
title_full | Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs |
title_fullStr | Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs |
title_full_unstemmed | Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs |
title_short | Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs |
title_sort | comparative study of real-time pcr (taqman probe and sybr green), serological techniques (elisa, ifa and dat) and clinical signs evaluation, for the diagnosis of canine leishmaniasis in experimentally infected dogs |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8704913/ https://www.ncbi.nlm.nih.gov/pubmed/34946227 http://dx.doi.org/10.3390/microorganisms9122627 |
work_keys_str_mv | AT perismariapaz comparativestudyofrealtimepcrtaqmanprobeandsybrgreenserologicaltechniqueselisaifaanddatandclinicalsignsevaluationforthediagnosisofcanineleishmaniasisinexperimentallyinfecteddogs AT estebangiladriana comparativestudyofrealtimepcrtaqmanprobeandsybrgreenserologicaltechniqueselisaifaanddatandclinicalsignsevaluationforthediagnosisofcanineleishmaniasisinexperimentallyinfecteddogs AT ortegahernandezpaula comparativestudyofrealtimepcrtaqmanprobeandsybrgreenserologicaltechniqueselisaifaanddatandclinicalsignsevaluationforthediagnosisofcanineleishmaniasisinexperimentallyinfecteddogs AT moralesmariano comparativestudyofrealtimepcrtaqmanprobeandsybrgreenserologicaltechniqueselisaifaanddatandclinicalsignsevaluationforthediagnosisofcanineleishmaniasisinexperimentallyinfecteddogs AT halaihelnabil comparativestudyofrealtimepcrtaqmanprobeandsybrgreenserologicaltechniqueselisaifaanddatandclinicalsignsevaluationforthediagnosisofcanineleishmaniasisinexperimentallyinfecteddogs AT castillojuanantonio comparativestudyofrealtimepcrtaqmanprobeandsybrgreenserologicaltechniqueselisaifaanddatandclinicalsignsevaluationforthediagnosisofcanineleishmaniasisinexperimentallyinfecteddogs |