4 results on '"Parrado, Rudy"'
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2. First external quality assurance program for bloodstream Real-Time PCR monitoring of treatment response in clinical trials of Chagas disease
- Author
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Ramírez, Juan C., Parrado, Rudy, Sulleiro Igual, Elena, de la Barra, Anabelle, Rodríguez, Marcelo, Villarroel, Sandro, Irazu, Lucía, Alonso-Vega, Cristina, Alves, Fabiana, Curto, María A., García, Lineth, Ortiz, Lourdes, Torrico, Faustino, Gascon, Joaquim, Flevaud, Laurence, Molina Romero, Israel, Ribeiro, Isabela, Schijman, Alejandro G., Universitat Autònoma de Barcelona, and Universitat de Barcelona
- Subjects
0301 basic medicine ,Chagas disease ,Posaconazole ,Molecular biology ,lcsh:Medicine ,Artificial Gene Amplification and Extension ,Satellite DNA ,Ravuconazole ,Polymerase Chain Reaction ,Biochemistry ,Geographical locations ,purl.org/becyt/ford/1 [https] ,0302 clinical medicine ,Malaltia de Chagas ,Medicine and Health Sciences ,lcsh:Science ,Protozoans ,Trypanosoma Cruzi ,Multidisciplinary ,External quality assurance ,Eukaryota ,Trypanocidal Agents ,Clinical Laboratory Sciences ,3. Good health ,Nucleic acids ,Clinical Laboratories ,Nitroimidazoles ,Benznidazole ,CIENCIAS NATURALES Y EXACTAS ,Research Article ,medicine.drug ,Trypanosoma ,Bolivia ,medicine.medical_specialty ,Drug Research and Development ,Forms of DNA ,Otras Ciencias Biológicas ,Concordance ,030231 tropical medicine ,030106 microbiology ,Real-Time Polymerase Chain Reaction ,Research and Analysis Methods ,Ciencias Biológicas ,03 medical and health sciences ,Diagnostic Medicine ,Internal medicine ,parasitic diseases ,Parasitic Diseases ,Genetics ,medicine ,Humans ,Chagas Disease ,Clinical Trials ,purl.org/becyt/ford/1.6 [https] ,Molecular Biology Techniques ,Molecular Biology ,Monitoring, Physiologic ,Biologia molecular ,Pharmacology ,proficiency testing ,business.industry ,lcsh:R ,Organisms ,Biology and Life Sciences ,DNA ,Triazoles ,South America ,Molecular diagnostics ,medicine.disease ,Parasitic Protozoans ,Clinical trial ,Chagas' disease ,Immunology ,lcsh:Q ,Real-Time PCR ,Clinical Medicine ,People and places ,business ,Quality assurance - Abstract
Real-Time PCR (qPCR) testing is recommended as both a diagnostic and outcome measurement of etiological treatment in clinical practice and clinical trials of Chagas disease (CD), but no external quality assurance (EQA) program provides performance assessment of the assays in use. We implemented an EQA system to evaluate the performance of molecular biology laboratories involved in qPCR based follow-up in clinical trials of CD. An EQA program was devised for three clinical trials of CD: the E1224 (NCT01489228), a pro-drug of ravuconazole; the Sampling Study (NCT01678599), that used benznidazole, both conducted in Bolivia; and the CHAGASAZOL (NCT01162967), that tested posaconazole, conducted in Spain. Four proficiency testing panels containing negative controls and seronegative blood samples spiked with 1, 10 and 100 parasite equivalents (par. eq.)/mL of four Trypanosoma cruzi stocks, were sent from the Core Lab in Argentina to the participating laboratories located in Bolivia and Spain. Panels were analyzed simultaneously, blinded to sample allocation, at 4-month intervals. In addition, 302 random blood samples from both trials carried out in Bolivia were sent to Core Lab for retesting analysis. The analysis of proficiency testing panels gave 100% of accordance (within laboratory agreement) and concordance (between laboratory agreement) for all T. cruzi stocks at 100 par. eq./mL; whereas their values ranged from 71 to 100% and from 62 to 100% at 1 and 10 par. eq./mL, respectively, depending on the T. cruzi stock. The results obtained after twelve months of preparation confirmed the stability of blood samples in guanidine-EDTA buffer. No significant differences were found between qPCR results from Bolivian laboratory and Core Lab for retested clinical samples. This EQA program for qPCR analysis of CD patient samples may significantly contribute to ensuring the quality of laboratory data generated in clinical trials and molecular diagnostics laboratories of CD. Fil: Ramirez Gomez, Juan Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: Parrado, Rudy. Universidad Mayor de San Simon; Bolivia Fil: Sulleiro, Elena. Universitat Autònoma de Barcelona; España Fil: De La Barra, Anabelle. Universidad Mayor de San Simon; Bolivia Fil: Rodríguez, Marcelo. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas; Argentina Fil: Villarroel, Sandro. Universidad Mayor de San Simon; Bolivia Fil: Irazu, Lucía. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas; Argentina Fil: Alonso Vega, Cristina. Drugs for Neglected Diseases initiative; Suiza Fil: Alves, Fabiana. Drugs for Neglected Diseases initiative; Suiza Fil: Curto, Maria de Los Angeles. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina Fil: García, Lineth. Universidad Mayor de San Simon; Bolivia Fil: Ortiz, Lourdes. Universidad Autonoma Juan Misael Saracho; Bolivia Fil: Torrico, Faustino. Fundacion CEADES; Bolivia Fil: Gascón, Joaquim. Universidad de Barcelona; España Fil: Flevaud, Laurence. Medecins Sans Frontières Operational Center Barcelona-Athens; España Fil: Molina, Israel. Universitat Autònoma de Barcelona; España Fil: Ribeiro, Isabela. Drugs for Neglected Diseases initiative; Suiza Fil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
- Published
- 2017
3. Sequence analysis of the 3'-untranslated region of HSP70 (type I) genes in the genus Leishmania: its usefulness as a molecular marker for species identification.
- Author
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Requena, Jose M, Chicharro, Carmen, Garc�a, Lineth, Parrado, Rudy, Puerta, Concepci�n J, and Ca�avate, Carmen
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LEISHMANIA ,LEISHMANIASIS ,PROGNOSIS ,PHYLOGENY ,POLYMERASE chain reaction - Abstract
Abstract: Background: The Leishmaniases are a group of clinically diverse diseases caused by parasites of the genus Leishmania. To distinguish between species is crucial for correct diagnosis and prognosis as well as for treatment decisions. Recently, sequencing of the HSP70 coding region has been applied in phylogenetic studies and for identifying of Leishmania species with excellent results. Methods: In the present study, we analyzed the 3'-untranslated region (UTR) of Leishmania HSP70-type I gene from 24 strains representing eleven Leishmania species in the belief that this non-coding region would have a better discriminatory capacity for species typing than coding regions. Results: It was observed that there was a remarkable degree of sequence conservation in this region, even between species of the subgenus Leishmania and Viannia. In addition, the presence of many microsatellites was a common feature of the 3´-UTR of HSP70-I genes in the Leishmania genus. Finally, we constructed dendrograms based on global sequence alignments of the analyzed Leishmania species and strains, the results indicated that this particular region of HSP70 genes might be useful for species (or species complex) typing, improving for particular species the discrimination capacity of phylogenetic trees based on HSP70 coding sequences. Given the large size variation of the analyzed region between the Leishmania and Viannia subgenera, direct visualization of the PCR amplification product would allow discrimination between subgenera, and a HaeIII-PCR-RFLP analysis might be used for differentiating some species within each subgenera. Conclusions: Sequence and phylogenetic analyses indicated that this region, which is readily amplified using a single pair of primers from both Old and New World Leishmania species, might be useful as a molecular marker for species discrimination. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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4. American tegumentary leishmaniasis: direct species identification of Leishmania in non-invasive clinical samples
- Author
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Garcia, Ana Lineth, Parrado, Rudy, De Doncker, Simonne, Bermudez, Hernán, and Dujardin, Jean-Claude
- Subjects
LEISHMANIASIS ,LEISHMANIA ,POLYMERASE chain reaction ,PATIENTS - Abstract
Summary: Species identification is highly relevant for improved prognosis and adequate treatment of American tegumentary leishmaniasis (ATL). PCR-based methods are available for this purpose but should be simplified to improve accessibility. As a first step in this process, this paper describes a simplified protocol for collection of clinical samples. Using samples from 44 Bolivian patients with confirmed ATL, we demonstrated that hsp70 PCR–RFLP on skin scrapings collected with a tooth pick allowed identification of the parasite species with a sensitivity of 95% and specificity of 100%. Our method should greatly facilitate individual patient management and epidemiological surveillance of ATL. [Copyright &y& Elsevier]
- Published
- 2007
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