International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients

ABSTRACT: Background: A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the...

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Autores:
Schijman, Alejandro
Bisio, Margarita
Orellana, Liliana
Sued, Mariela
Duffy, Tomás
Mejía Jaramillo, Ana María
Cura, Carolina
Auter, Frederic
Veron, Vincent
Qvarnstrom, Yvonne
Deborggraeve, Stijn
Hijar, Gisely
Zulantay, Inés
Lucero, Raúl Horacio
Velázquez, Elsa
Tellez, Tatiana
Sánchez León, Zunilda
Galvão, Lucia
Nolder, Debbie
Monje Rumi, María
Levi, José
Ramírez, Juan
Zorrilla, Pilar
Flores, María
Jercic, Maria
Crisante, Gladys
Añez, Néstor
de Castro, Ana
González, Clara
Acosta Viana, Karla
Yachelini, Pedro
Torrico, Faustino
Robello, Carlos
Diosque, Patricio
Triana Chávez, Omar
Aznar, Christine
Russomando, Graciela
Büscher, Philippe
Assal, Azzedine
Guhl, Felipe
Sosa Estani, Sergio
DaSilva, Alexandre
Britto, Constança
Luquetti, Alejandro
Ladzins, Janis
Tipo de recurso:
Article of investigation
Fecha de publicación:
2011
Institución:
Universidad de Antioquia
Repositorio:
Repositorio UdeA
Idioma:
eng
OAI Identifier:
oai:bibliotecadigital.udea.edu.co:10495/23712
Acceso en línea:
http://hdl.handle.net/10495/23712
Palabra clave:
Enfermedad de chagas
Chagas Disease
Trypanosoma cruzi
Reacción en Cadena de la Polimerasa
Polymerase Chain Reaction
Rights
openAccess
License
http://creativecommons.org/licenses/by/2.5/co/
id UDEA2_09df24b5e46640f1a9914be404cb43a3
oai_identifier_str oai:bibliotecadigital.udea.edu.co:10495/23712
network_acronym_str UDEA2
network_name_str Repositorio UdeA
repository_id_str
dc.title.spa.fl_str_mv International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
title International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
spellingShingle International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
Enfermedad de chagas
Chagas Disease
Trypanosoma cruzi
Reacción en Cadena de la Polimerasa
Polymerase Chain Reaction
title_short International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
title_full International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
title_fullStr International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
title_full_unstemmed International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
title_sort International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients
dc.creator.fl_str_mv Schijman, Alejandro
Bisio, Margarita
Orellana, Liliana
Sued, Mariela
Duffy, Tomás
Mejía Jaramillo, Ana María
Cura, Carolina
Auter, Frederic
Veron, Vincent
Qvarnstrom, Yvonne
Deborggraeve, Stijn
Hijar, Gisely
Zulantay, Inés
Lucero, Raúl Horacio
Velázquez, Elsa
Tellez, Tatiana
Sánchez León, Zunilda
Galvão, Lucia
Nolder, Debbie
Monje Rumi, María
Levi, José
Ramírez, Juan
Zorrilla, Pilar
Flores, María
Jercic, Maria
Crisante, Gladys
Añez, Néstor
de Castro, Ana
González, Clara
Acosta Viana, Karla
Yachelini, Pedro
Torrico, Faustino
Robello, Carlos
Diosque, Patricio
Triana Chávez, Omar
Aznar, Christine
Russomando, Graciela
Büscher, Philippe
Assal, Azzedine
Guhl, Felipe
Sosa Estani, Sergio
DaSilva, Alexandre
Britto, Constança
Luquetti, Alejandro
Ladzins, Janis
dc.contributor.author.none.fl_str_mv Schijman, Alejandro
Bisio, Margarita
Orellana, Liliana
Sued, Mariela
Duffy, Tomás
Mejía Jaramillo, Ana María
Cura, Carolina
Auter, Frederic
Veron, Vincent
Qvarnstrom, Yvonne
Deborggraeve, Stijn
Hijar, Gisely
Zulantay, Inés
Lucero, Raúl Horacio
Velázquez, Elsa
Tellez, Tatiana
Sánchez León, Zunilda
Galvão, Lucia
Nolder, Debbie
Monje Rumi, María
Levi, José
Ramírez, Juan
Zorrilla, Pilar
Flores, María
Jercic, Maria
Crisante, Gladys
Añez, Néstor
de Castro, Ana
González, Clara
Acosta Viana, Karla
Yachelini, Pedro
Torrico, Faustino
Robello, Carlos
Diosque, Patricio
Triana Chávez, Omar
Aznar, Christine
Russomando, Graciela
Büscher, Philippe
Assal, Azzedine
Guhl, Felipe
Sosa Estani, Sergio
DaSilva, Alexandre
Britto, Constança
Luquetti, Alejandro
Ladzins, Janis
dc.subject.decs.none.fl_str_mv Enfermedad de chagas
Chagas Disease
Trypanosoma cruzi
Reacción en Cadena de la Polimerasa
Polymerase Chain Reaction
topic Enfermedad de chagas
Chagas Disease
Trypanosoma cruzi
Reacción en Cadena de la Polimerasa
Polymerase Chain Reaction
description ABSTRACT: Background: A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the performance of PCR methods in detection of Trypanosoma cruzi DNA by an external quality evaluation. Methodology/Findings: An international collaborative study was launched by expert PCR laboratories from 16 countries. Currently used strategies were challenged against serial dilutions of purified DNA from stocks representing T. cruzi discrete typing units (DTU) I, IV and VI (set A), human blood spiked with parasite cells (set B) and Guanidine Hidrochloride-EDTA blood samples from 32 seropositive and 10 seronegative patients from Southern Cone countries (set C). Forty eight PCR tests were reported for set A and 44 for sets B and C; 28 targeted minicircle DNA (kDNA), 13 satellite DNA (Sat-DNA) and the remainder low copy number sequences. In set A, commercial master mixes and Sat-DNA Real Time PCR showed better specificity, but kDNA-PCR was more sensitive to detect DTU I DNA. In set B, commercial DNA extraction kits presented better specificity than solvent extraction protocols. Sat-DNA PCR tests had higher specificity, with sensitivities of 0.05–0.5 parasites/ mL whereas specific kDNA tests detected 5.1023 par/mL. Sixteen specific and coherent methods had a Good Performance in both sets A and B (10 fg/ml of DNA from all stocks, 5 par/mL spiked blood). The median values of sensitivities, specificities and accuracies obtained in testing the Set C samples with the 16 tests determined to be good performing by analyzing Sets A and B samples varied considerably. Out of them, four methods depicted the best performing parameters in all three sets of samples, detecting at least 10 fg/ml for each DNA stock, 0.5 par/mL and a sensitivity between 83.3–94.4%, specificity of 85–95%, accuracy of 86.8–89.5% and kappa index of 0.7–0.8 compared to consensus PCR reports of the 16 good performing tests and 63–69%, 100%, 71.4–76.2% and 0.4–0.5, respectively compared to serodiagnosis. Method LbD2 used solvent extraction followed by Sybr-Green based Real time PCR targeted to Sat-DNA; method LbD3 used solvent DNA extraction followed by conventional PCR targeted to Sat-DNA. The third method (LbF1) used glass fiber column based DNA extraction followed by TaqMan Real Time PCR targeted to Sat-DNA (cruzi 1/cruzi 2 and cruzi 3 TaqMan probe) and the fourth method (LbQ) used solvent DNA extraction followed by conventional hot-start PCR targeted to kDNA (primer pairs 121/122). These four methods were further evaluated at the coordinating laboratory in a subset of human blood samples, confirming the performance obtained by the participating laboratories. Conclusion/Significance: This study represents a first crucial step towards international validation of PCR procedures for detection of T. cruzi in human blood samples.
publishDate 2011
dc.date.issued.none.fl_str_mv 2011
dc.date.accessioned.none.fl_str_mv 2021-11-02T22:42:12Z
dc.date.available.none.fl_str_mv 2021-11-02T22:42:12Z
dc.type.spa.fl_str_mv info:eu-repo/semantics/article
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dc.type.local.spa.fl_str_mv Artículo de investigación
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dc.identifier.citation.spa.fl_str_mv Schijman AG, Bisio M, Orellana L, Sued M, Duffy T, et al. (2011) International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. PLoS Negl Trop Dis 5(1): e931. doi:10.1371/journal.pntd.0000931
dc.identifier.issn.none.fl_str_mv 1553-7390
dc.identifier.uri.none.fl_str_mv http://hdl.handle.net/10495/23712
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pntd.0000931
dc.identifier.eissn.none.fl_str_mv 1553-7404
identifier_str_mv Schijman AG, Bisio M, Orellana L, Sued M, Duffy T, et al. (2011) International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. PLoS Negl Trop Dis 5(1): e931. doi:10.1371/journal.pntd.0000931
1553-7390
10.1371/journal.pntd.0000931
1553-7404
url http://hdl.handle.net/10495/23712
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.ispartofjournalabbrev.spa.fl_str_mv PLoS Negl. Trop. Dis.
dc.rights.spa.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.spa.fl_str_mv Public Library of Science
dc.publisher.group.spa.fl_str_mv Genética Molecular (GENMOL)
dc.publisher.place.spa.fl_str_mv San Francisco, Estados Unidos
institution Universidad de Antioquia
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spelling Schijman, AlejandroBisio, MargaritaOrellana, LilianaSued, MarielaDuffy, TomásMejía Jaramillo, Ana MaríaCura, CarolinaAuter, FredericVeron, VincentQvarnstrom, YvonneDeborggraeve, StijnHijar, GiselyZulantay, InésLucero, Raúl HoracioVelázquez, ElsaTellez, TatianaSánchez León, ZunildaGalvão, LuciaNolder, DebbieMonje Rumi, MaríaLevi, JoséRamírez, JuanZorrilla, PilarFlores, MaríaJercic, MariaCrisante, GladysAñez, Néstorde Castro, AnaGonzález, ClaraAcosta Viana, KarlaYachelini, PedroTorrico, FaustinoRobello, CarlosDiosque, PatricioTriana Chávez, OmarAznar, ChristineRussomando, GracielaBüscher, PhilippeAssal, AzzedineGuhl, FelipeSosa Estani, SergioDaSilva, AlexandreBritto, ConstançaLuquetti, AlejandroLadzins, Janis2021-11-02T22:42:12Z2021-11-02T22:42:12Z2011Schijman AG, Bisio M, Orellana L, Sued M, Duffy T, et al. (2011) International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients. PLoS Negl Trop Dis 5(1): e931. doi:10.1371/journal.pntd.00009311553-7390http://hdl.handle.net/10495/2371210.1371/journal.pntd.00009311553-7404ABSTRACT: Background: A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the performance of PCR methods in detection of Trypanosoma cruzi DNA by an external quality evaluation. Methodology/Findings: An international collaborative study was launched by expert PCR laboratories from 16 countries. Currently used strategies were challenged against serial dilutions of purified DNA from stocks representing T. cruzi discrete typing units (DTU) I, IV and VI (set A), human blood spiked with parasite cells (set B) and Guanidine Hidrochloride-EDTA blood samples from 32 seropositive and 10 seronegative patients from Southern Cone countries (set C). Forty eight PCR tests were reported for set A and 44 for sets B and C; 28 targeted minicircle DNA (kDNA), 13 satellite DNA (Sat-DNA) and the remainder low copy number sequences. In set A, commercial master mixes and Sat-DNA Real Time PCR showed better specificity, but kDNA-PCR was more sensitive to detect DTU I DNA. In set B, commercial DNA extraction kits presented better specificity than solvent extraction protocols. Sat-DNA PCR tests had higher specificity, with sensitivities of 0.05–0.5 parasites/ mL whereas specific kDNA tests detected 5.1023 par/mL. Sixteen specific and coherent methods had a Good Performance in both sets A and B (10 fg/ml of DNA from all stocks, 5 par/mL spiked blood). The median values of sensitivities, specificities and accuracies obtained in testing the Set C samples with the 16 tests determined to be good performing by analyzing Sets A and B samples varied considerably. Out of them, four methods depicted the best performing parameters in all three sets of samples, detecting at least 10 fg/ml for each DNA stock, 0.5 par/mL and a sensitivity between 83.3–94.4%, specificity of 85–95%, accuracy of 86.8–89.5% and kappa index of 0.7–0.8 compared to consensus PCR reports of the 16 good performing tests and 63–69%, 100%, 71.4–76.2% and 0.4–0.5, respectively compared to serodiagnosis. Method LbD2 used solvent extraction followed by Sybr-Green based Real time PCR targeted to Sat-DNA; method LbD3 used solvent DNA extraction followed by conventional PCR targeted to Sat-DNA. The third method (LbF1) used glass fiber column based DNA extraction followed by TaqMan Real Time PCR targeted to Sat-DNA (cruzi 1/cruzi 2 and cruzi 3 TaqMan probe) and the fourth method (LbQ) used solvent DNA extraction followed by conventional hot-start PCR targeted to kDNA (primer pairs 121/122). These four methods were further evaluated at the coordinating laboratory in a subset of human blood samples, confirming the performance obtained by the participating laboratories. Conclusion/Significance: This study represents a first crucial step towards international validation of PCR procedures for detection of T. cruzi in human blood samples.COL000672313application/pdfengPublic Library of ScienceGenética Molecular (GENMOL)San Francisco, Estados Unidosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARTArtículo de investigaciónhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by/4.0/International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease PatientsEnfermedad de chagasChagas DiseaseTrypanosoma cruziReacción en Cadena de la PolimerasaPolymerase Chain ReactionPLoS Negl. Trop. Dis.PLoS Neglected Tropical Diseases11351ORIGINALJaramilloAna_2011_StudyDetectionTrypanosoma.pdfJaramilloAna_2011_StudyDetectionTrypanosoma.pdfArtículo de investigaciónapplication/pdf885168http://bibliotecadigital.udea.edu.co/bitstream/10495/23712/1/JaramilloAna_2011_StudyDetectionTrypanosoma.pdf899a7d486740a706f8e5aaeb05fb7dc4MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8927http://bibliotecadigital.udea.edu.co/bitstream/10495/23712/2/license_rdf1646d1f6b96dbbbc38035efc9239ac9cMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://bibliotecadigital.udea.edu.co/bitstream/10495/23712/3/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5310495/23712oai:bibliotecadigital.udea.edu.co:10495/237122021-11-02 17:42:13.027Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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