Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients

Aim: Here, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection of Leishmania infection in Cutaneous Leishmaniasis patients. Methods: A standard curve of cultured Leishmania parasite material and clinical samples of CL patients were tested with ddPCR to det...

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Autores:
Tipo de recurso:
Fecha de publicación:
2019
Institución:
Universidad del Rosario
Repositorio:
Repositorio EdocUR - U. Rosario
Idioma:
eng
OAI Identifier:
oai:repository.urosario.edu.co:10336/22126
Acceso en línea:
https://doi.org/10.1016/j.ijid.2018.10.029
https://repository.urosario.edu.co/handle/10336/22126
Palabra clave:
Protozoal DNA
Protozoal DNA
Article
Droplet digital polymerase chain reaction
Human
Leishmania
Leishmania braziliensis
Leishmania donovani
Leishmania guyanensis
Leishmania infantum
Leishmania major
Leishmania mexicana
Leishmania panamensis
Limit of detection
Microorganism detection
Molecular pathology
Skin leishmaniasis
Task performance
Validation study
Evaluation study
Female
Genetics
Isolation and purification
Leishmania
Parasitology
Procedures
Real time polymerase chain reaction
Sensitivity and specificity
Skin leishmaniasis
Female
Humans
Leishmania
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Cutaneous Leishmaniasis
Ddpcr
Leishmania
Molecular diagnosis
Cutaneous
Protozoan
DNA
Leishmaniasis
Rights
License
Abierto (Texto Completo)
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spelling 10117161186005193546460007f5f951-ff31-4b0f-94f0-7492d0a86f05-194094a23-a4e4-4418-b743-dac06fb0417f-1a34b10e4-f719-4e6a-9d4f-0717561679cc-181002983-a1d5-453d-983d-72b289a34d02-12020-05-25T23:55:33Z2020-05-25T23:55:33Z2019Aim: Here, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection of Leishmania infection in Cutaneous Leishmaniasis patients. Methods: A standard curve of cultured Leishmania parasite material and clinical samples of CL patients were tested with ddPCR to determine the analytical and diagnostic performance. Results: The limit of detection of the assay on the ddPCR platform was much higher than the published limit of detection of the same assay on the qPCR platform (100 vs 1 parasites/mL, respectively). Conclusion: While the performance of this assay in ddPCR format was acceptable for research purposes, it is not sufficient for clinical diagnostic purposes. The assay is more suited to the qPCR platform. © 2018 The Author(s)application/pdfhttps://doi.org/10.1016/j.ijid.2018.10.02912019712https://repository.urosario.edu.co/handle/10336/22126engElsevier B.V.31International Journal of Infectious DiseasesVol. 79International Journal of Infectious Diseases, ISSN:12019712, Vol.79,(2019); pp. 1-3https://www.scopus.com/inward/record.uri?eid=2-s2.0-85057876716&doi=10.1016%2fj.ijid.2018.10.029&partnerID=40&md5=df03529c552cdaa5301dfc4023ee8107Abierto (Texto Completo)http://purl.org/coar/access_right/c_abf2instname:Universidad del Rosarioreponame:Repositorio Institucional EdocURProtozoal DNAProtozoal DNAArticleDroplet digital polymerase chain reactionHumanLeishmaniaLeishmania braziliensisLeishmania donovaniLeishmania guyanensisLeishmania infantumLeishmania majorLeishmania mexicanaLeishmania panamensisLimit of detectionMicroorganism detectionMolecular pathologySkin leishmaniasisTask performanceValidation studyEvaluation studyFemaleGeneticsIsolation and purificationLeishmaniaParasitologyProceduresReal time polymerase chain reactionSensitivity and specificitySkin leishmaniasisFemaleHumansLeishmaniaReal-Time Polymerase Chain ReactionSensitivity and SpecificityCutaneous LeishmaniasisDdpcrLeishmaniaMolecular diagnosisCutaneousProtozoanDNALeishmaniasisDevelopment of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patientsarticleArtículohttp://purl.org/coar/version/c_970fb48d4fbd8a85http://purl.org/coar/resource_type/c_6501Ramírez, Juan DavidMéndez-Sarmiento, Claudia Herrera, GiovannyMuskus, CarlosDuque, María ClaraButcher, RobertORIGINALPIIS1201971218345776.pdfapplication/pdf548343https://repository.urosario.edu.co/bitstreams/88b1052a-fdbd-48f4-8be2-96ea78659117/download1a9be378802862d647afee4f5cab3361MD51TEXTPIIS1201971218345776.pdf.txtPIIS1201971218345776.pdf.txtExtracted texttext/plain11046https://repository.urosario.edu.co/bitstreams/a7e02a74-acac-4dd3-a0a3-9a1bf5b75ac0/download3e533cb2f0f25923c9068805b57011feMD52THUMBNAILPIIS1201971218345776.pdf.jpgPIIS1201971218345776.pdf.jpgGenerated Thumbnailimage/jpeg4934https://repository.urosario.edu.co/bitstreams/66b7129f-91ac-46d2-bcea-761a7b078cdc/download42ad98ef0779ded1c993f54d03932d70MD5310336/22126oai:repository.urosario.edu.co:10336/221262022-05-02 07:37:18.066892https://repository.urosario.edu.coRepositorio institucional EdocURedocur@urosario.edu.co
dc.title.spa.fl_str_mv Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
title Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
spellingShingle Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
Protozoal DNA
Protozoal DNA
Article
Droplet digital polymerase chain reaction
Human
Leishmania
Leishmania braziliensis
Leishmania donovani
Leishmania guyanensis
Leishmania infantum
Leishmania major
Leishmania mexicana
Leishmania panamensis
Limit of detection
Microorganism detection
Molecular pathology
Skin leishmaniasis
Task performance
Validation study
Evaluation study
Female
Genetics
Isolation and purification
Leishmania
Parasitology
Procedures
Real time polymerase chain reaction
Sensitivity and specificity
Skin leishmaniasis
Female
Humans
Leishmania
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Cutaneous Leishmaniasis
Ddpcr
Leishmania
Molecular diagnosis
Cutaneous
Protozoan
DNA
Leishmaniasis
title_short Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
title_full Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
title_fullStr Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
title_full_unstemmed Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
title_sort Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients
dc.subject.keyword.spa.fl_str_mv Protozoal DNA
Protozoal DNA
Article
Droplet digital polymerase chain reaction
Human
Leishmania
Leishmania braziliensis
Leishmania donovani
Leishmania guyanensis
Leishmania infantum
Leishmania major
Leishmania mexicana
Leishmania panamensis
Limit of detection
Microorganism detection
Molecular pathology
Skin leishmaniasis
Task performance
Validation study
Evaluation study
Female
Genetics
Isolation and purification
Leishmania
Parasitology
Procedures
Real time polymerase chain reaction
Sensitivity and specificity
Skin leishmaniasis
Female
Humans
Leishmania
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Cutaneous Leishmaniasis
Ddpcr
Leishmania
Molecular diagnosis
topic Protozoal DNA
Protozoal DNA
Article
Droplet digital polymerase chain reaction
Human
Leishmania
Leishmania braziliensis
Leishmania donovani
Leishmania guyanensis
Leishmania infantum
Leishmania major
Leishmania mexicana
Leishmania panamensis
Limit of detection
Microorganism detection
Molecular pathology
Skin leishmaniasis
Task performance
Validation study
Evaluation study
Female
Genetics
Isolation and purification
Leishmania
Parasitology
Procedures
Real time polymerase chain reaction
Sensitivity and specificity
Skin leishmaniasis
Female
Humans
Leishmania
Real-Time Polymerase Chain Reaction
Sensitivity and Specificity
Cutaneous Leishmaniasis
Ddpcr
Leishmania
Molecular diagnosis
Cutaneous
Protozoan
DNA
Leishmaniasis
dc.subject.keyword.eng.fl_str_mv Cutaneous
Protozoan
DNA
Leishmaniasis
description Aim: Here, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection of Leishmania infection in Cutaneous Leishmaniasis patients. Methods: A standard curve of cultured Leishmania parasite material and clinical samples of CL patients were tested with ddPCR to determine the analytical and diagnostic performance. Results: The limit of detection of the assay on the ddPCR platform was much higher than the published limit of detection of the same assay on the qPCR platform (100 vs 1 parasites/mL, respectively). Conclusion: While the performance of this assay in ddPCR format was acceptable for research purposes, it is not sufficient for clinical diagnostic purposes. The assay is more suited to the qPCR platform. © 2018 The Author(s)
publishDate 2019
dc.date.created.spa.fl_str_mv 2019
dc.date.accessioned.none.fl_str_mv 2020-05-25T23:55:33Z
dc.date.available.none.fl_str_mv 2020-05-25T23:55:33Z
dc.type.eng.fl_str_mv article
dc.type.coarversion.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.coar.fl_str_mv http://purl.org/coar/resource_type/c_6501
dc.type.spa.spa.fl_str_mv Artículo
dc.identifier.doi.none.fl_str_mv https://doi.org/10.1016/j.ijid.2018.10.029
dc.identifier.issn.none.fl_str_mv 12019712
dc.identifier.uri.none.fl_str_mv https://repository.urosario.edu.co/handle/10336/22126
url https://doi.org/10.1016/j.ijid.2018.10.029
https://repository.urosario.edu.co/handle/10336/22126
identifier_str_mv 12019712
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.citationEndPage.none.fl_str_mv 3
dc.relation.citationStartPage.none.fl_str_mv 1
dc.relation.citationTitle.none.fl_str_mv International Journal of Infectious Diseases
dc.relation.citationVolume.none.fl_str_mv Vol. 79
dc.relation.ispartof.spa.fl_str_mv International Journal of Infectious Diseases, ISSN:12019712, Vol.79,(2019); pp. 1-3
dc.relation.uri.spa.fl_str_mv https://www.scopus.com/inward/record.uri?eid=2-s2.0-85057876716&doi=10.1016%2fj.ijid.2018.10.029&partnerID=40&md5=df03529c552cdaa5301dfc4023ee8107
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dc.rights.acceso.spa.fl_str_mv Abierto (Texto Completo)
rights_invalid_str_mv Abierto (Texto Completo)
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dc.publisher.spa.fl_str_mv Elsevier B.V.
institution Universidad del Rosario
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