International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients
| Autor: | Janis Ladzins, Gladys Crisante, Ana M. Mejia Jaramillo, Maria I. Jercic, Tatiana Tellez, Alexandre J. DaSilva, Stijn Deborggraeve, Alejandro O. Luquetti, Debbie Nolder, Pilar Zorrilla, Patricio Diosque, Maria Mercedes Monje Rumi, Lúcia Maria da Cunha Galvão, Sergio Sosa Estani, Carlos Robello, Omar Triana Chávez, Faustino Torrico, María Flores, Alejandro G. Schijman, Juan David Ramírez, Néstor Añez, Pedro Yachelini, Mariela Sued, Azzedine Assal, Raul Horacio Lucero, Constança Britto, Inés Zulantay, Gisely Hijar, Christine Aznar, Vincent Veron, Margarita Bisio, Carolina Cura, Ana Maria de Castro, Philippe Büscher, Elsa F. Velazquez, Tomás Duffy, Clara Isabel González, José Eduardo Levi, Frederic Auter, Felipe Guhl, Karla Y. Acosta Viana, Yvonne Qvarnstrom, Graciela Russomando, Liliana Orellana, Zunilda Sanchez Leon |
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| Přispěvatelé: | Laboratorio de Biologia Molecular de la Enfermedad de Chagas, Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Instituto de Cálculo [Buenos Aires], Facultad de Ciencias Exactas y Naturales [Buenos Aires] (FCEyN), Universidad de Buenos Aires [Buenos Aires] (UBA)-Universidad de Buenos Aires [Buenos Aires] (UBA), Grupo Chagas, Universidad de Antioquia = University of Antioquia [Medellín, Colombia], French Blood Services, Coordination Régionale de la Lutte contre le VIH, Centre Hospitalier Andrée Rosemon [Cayenne, Guyane Française], Department of parasitic diseases, Centers for Disease Control, Institute of Tropical Medicine [Antwerp] (ITM), Instituto nacional de Salud, Facultad de Medicina, Universidad Nacional del Nordeste [Corrientes] (UNNE), Instituto Nacional de Chagas, Instituto Nacional de Parasitología 'Dr. Mario Fatala Chaben' (INP), Centro universitario de Medicina Tropical, Facultad de Medicina-Universidad Mayor de San Simón [Cochabamba, Bolivie] (UMSS), Instituto de Investigaciones en Ciencias de la Salud, Universidad Nacional de Asunción [Paraguay] (UNA), Faculdade de Farmacia, Faculdade de Farmácia, Department of Clinical Parasitology, Hospital for Tropical Diseases-London School of Hygiene and Tropical Medicine (LSHTM), Instituto de Patología Experimental [Salta], Consejo Nacional de Investigaciones Científicas y Técnicas [Buenos Aires] (CONICET)-Facultad de Ciencias de la Salud [Salta], Universidad Nacional de Salta (UNSA)-Universidad Nacional de Salta (UNSA), Blood Bank, Hospital Sirio Libanes, Centro de Investigaciones en Microbiologia y Parasitologia Tropical, Universidad de los Andes [Bogota] (UNIANDES), Institut Pasteur de Montevideo, Réseau International des Instituts Pasteur (RIIP), Centro de Mahahonda, Instituto de Salud Carlos III [Madrid] (ISC)-Centro Nacional de Microbiologia, Seccion Parasitologia, Centro de Investigaciones Parasitologicas 'J.F Torrealba', Instituto de Patologia Tropical e Saúde Pública (IPTSP), Universidade Federal de Goiás [Goiânia] (UFG), Grupo de Inmunologia y Epidemiologia Molecular, Universidad Industrial de Santander [Bucaramanga] (UIS)-Facultad de Salud, Departamento de Biomedicina de Enfermedades Infecciosas y Parasitarias, Laboratorio de Biologia Celular, Centro de Investigaciones Regionales 'Dr Hideyo Noguchi'-Universidad Autónoma de Yucatán, Instituto de Biomedicina, Universidad Catolica de Santiago del Estero, Centro Nacional de Diagnostico e Investigacion de Endemoepidemias, Laboratory of Molecular Biology and Diagnosis of Infectious Diseases / Laboratório de Biologia Molecular e Doenças Endêmicas [Rio de Janeiro], Instituto Oswaldo Cruz / Oswaldo Cruz Institute [Rio de Janeiro] (IOC), Fundação Oswaldo Cruz (FIOCRUZ), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Fundação Oswaldo Cruz (FIOCRUZ), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Laboratorio de pesquisa de Doença de Chagas, Special Programme for Research and Training in Tropical Diseases, Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Funding was provided by World Health Organization-Tropical Diseases Research, Panamerican Health Organization (WHO-TDR-PAHO), Universidad de las Naciones Unidas/Biotecnologia para America Latina y el Caribe (UNU-BIOLAC), Consejo Nacional de Ciencias y Tecnologia (CONICET) PIP 112-200801-09215, National Agency of Science and Technology, PICT 33955, Buenos Aires, Argentina., World Health Organization (WHO/OMS), Organización Panamericana de la Salud, United Nations University, Consejo Nacional de Investigaciones Científicas y Técnicas (Argentina) |
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
lcsh:Arctic medicine. Tropical medicine
Enfermedad de chagas Satellite DNA lcsh:RC955-962 International Cooperation Trypanosoma cruzi 030231 tropical medicine Consensus PCR MESH: DNA Protozoan MESH: Parasitology Sensitivity and Specificity Polymerase Chain Reaction law.invention 03 medical and health sciences 0302 clinical medicine law TaqMan Humans Chagas Disease MESH: Chagas Disease Microbiology/Parasitology Molecular Biology Polymerase chain reaction 0303 health sciences [SDV.GEN]Life Sciences [q-bio]/Genetics MESH: Humans biology 030306 microbiology lcsh:Public aspects of medicine Infectious Diseases/Protozoal Infections Public Health Environmental and Occupational Health MESH: Polymerase Chain Reaction lcsh:RA1-1270 DNA Protozoan biology.organism_classification DNA extraction Molecular biology MESH: Sensitivity and Specificity 3. Good health MESH: International Cooperation Infectious Diseases Real-time polymerase chain reaction Infectious Diseases/Neglected Tropical Diseases Reacción en Cadena de la Polimerasa Parasitology Low copy number MESH: Trypanosoma cruzi Research Article |
| Zdroj: | PLoS Neglected Tropical Diseases, Vol 5, Iss 1, p e931 (2011) PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Public Library of Science, 2011, 5 (1), pp.e931. ⟨10.1371/journal.pntd.0000931⟩ Repositorio UdeA Universidad de Antioquia instacron:Universidad de Antioquia Repisalud Instituto de Salud Carlos III (ISCIII) PLoS Neglected Tropical Diseases; Vol 5 |
| ISSN: | 1935-2735 1935-2727 |
| DOI: | 10.1371/journal.pntd.0000931⟩ |
| Popis: | 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 5.10−3 par/mL. Sixteen specific and coherent methods had a Good Performance in both sets A and B (10 fg/µl 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/µl 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. Author Summary A century after its discovery, Chagas disease, caused by the parasite Trypanosoma cruzi, 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 polymerase chain reaction (PCR) has been proposed as a sensitive laboratory tool for detection of T. cruzi infection and monitoring of parasitological treatment outcome. However, high variation in accuracy and lack of international quality controls has precluded reliable applications in the clinical practice and comparisons of data among cohorts and geographical regions. In an effort towards harmonization of PCR strategies, 26 expert laboratories from 16 countries evaluated their current PCR procedures against sets of control samples, composed by serial dilutions of T.cruzi DNA from culture stocks belonging to different lineages, human blood spiked with parasite cells and blood samples from Chagas disease patients. A high variability in sensitivities and specificities was found among the 48 reported PCR tests. Out of them, four tests with best performance were selected and further evaluated. This study represents a crucial first step towards device of a standardized operative procedure for T. cruzi PCR. |
| Databáze: | OpenAIRE |
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