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Featured researches published by Sébastien Boucher.


Malaria Journal | 2008

Evaluation of FRET real-time PCR assay for rapid detection and differentiation of Plasmodium species in returning travellers and migrants

Innocent Safeukui; Pascal Millet; Sébastien Boucher; Laurence Melinard; Frédéric Fregeville; Marie-Catherine Receveur; Thierry Pistone; Pierre Fialon; Philippe Vincendeau; Hervé Fleury; Denis Malvy

BackgroundA simple real-time PCR assay using one set of primer and probe for rapid, sensitive and quantitative detection of Plasmodium species, with simultaneous differentiation of Plasmodium falciparum from the three other Plasmodium species (Plasmodium vivax, Plasmodium ovale and Plasmodium malariae) in febrile returning travellers and migrants was developed and evaluated.MethodsConsensus primers were used to amplify a species-specific region of the multicopy 18S rRNA gene, and fluorescence resonance energy transfer hybridization probes were used for detection in a LightCycler platform (Roche). The anchor probe sequence was designed to be perfect matches to the 18S rRNA gene of the fourth Plasmodium species, while the acceptor probe sequence was designed for P. falciparum over a region containing one mismatched, which allowed differentiation of the three other Plasmodium species. The performance characteristics of the real-time PCR assay were compared with those of conventional PCR and microscopy-based diagnosis from 119 individuals with a suspected clinical diagnostic of imported malaria.ResultsBlood samples with parasite densities less than 0.01% were all detected, and analytical sensitivity was 0.5 parasite per PCR reaction. The melt curve means Tms (standard deviation) in clinical isolates were 60.5°C (0.6°C) for P. falciparum infection and 64.6°C (1.8°C) for non-P. falciparum species. These Tms values of the P. falciparum or non-P. falciparum species did not vary with the geographic origin of the parasite. The real-time PCR results correlated with conventional PCR using both genus-specific (Kappa coefficient: 0.95, 95% confidence interval: 0.9 – 1) or P. falciparum-specific (0.91, 0.8 – 1) primers, or with the microscopy results (0.70, 0.6 – 0.8). The real-time assay was 100% sensitive and specific for differentiation of P. falciparum to non-P. falciparum species, compared with conventional PCR or microscopy. The real-time PCR assay can also detect individuals with mixed infections (P. falciparum and non-P. falciparum sp.) in the same sample.ConclusionThis real-time PCR assay with melting curve analysis is rapid, and specific for the detection and differentiation of P. falciparum to other Plasmodium species. The suitability for routine use of this assay in clinical diagnostic laboratories is discussed.


Antimicrobial Agents and Chemotherapy | 2007

Interpretation of Genotype and Pharmacokinetics for Resistance to Fosamprenavir-Ritonavir-Based Regimens in Antiretroviral-Experienced Patients

Isabelle Pellegrin; Dominique Breilh; Gaëlle Coureau; Sébastien Boucher; Didier Neau; Patrick Merel; Denis Lacoste; Hervé Fleury; Marie-Claude Saux; Jean-Luc Pellegrin; Estibaliz Lazaro; François Dabis; Rodolphe Thiébaut

ABSTRACT In this study, named the Zephir study (Telzir-pharmacokinetics), 121 antiretroviral-experienced human immunodeficiency virus (HIV) patients failing on highly active antiretroviral therapy (HAART) were included in a prospective cohort and received a fosamprenavir-ritonavir (700 mg/100 mg twice a day)-based regimen. The impact of baseline HIV type 1 (HIV-1) mutations, pharmacokinetic (PK) parameters, and genotype inhibitory quotient (GIQ) on the virological response at week 12 (W12) was assessed. HIV reverse transcriptase and protease were sequenced at W0. The response at W12 was defined as <2.3 log10 HIV-1 RNA copies/ml or a virus load decrease of ≥1 log10 copies/ml. W4 amprenavir PK were determined by high-performance liquid chromatography. Patients had a median of nine previous treatments over 8 years. Median W0 values were as follows: 295 CD4+/μl, 4.4 log10 HIV-1 RNA copies/ml, and 6 protease- and 5 nucleotide reverse transcription inhibitor-related mutations. Respective values for minimum concentration of drug in serum (Cmin) and area under the concentration-time curve (AUC) from 0 to 24 h were 1,400 ng/ml and 35 mg·h/ml. At W12, 52% of the patients were successes, with a median decrease of −0.7 log10 HIV-1 RNA copies/ml. The Zephir mutation score included 12 IAS protease mutations associated with poorer virological response: L10I/F/R/V, L33F, M36I, M46I/L, I54L/M/T/V, I62V, L63P, A71I/L/V/T, G73A/C/F/T, V82A/F/S/T, I84V, L90M, and polymorphism mutations I13V, L19I, K55R, and L89M. Comparing <4 versus ≥4 mutations, HIV-1 RNA decreases were −2.3 log10 copies/ml versus −0.1 log10 copies/ml (P < 10−4) with 93% versus 19% successes (P < 10−4), respectively. This score predicted W12 failure with 94% sensitivity, versus 31% for the ANRS 2005 algorithm. Cmin (<1,600 ng/ml), AUC (<40 mg·h/ml), and GIQ (<300) values were associated with failure (all P values were <10−4). The need to test genotype-based algorithms using different patient databases before their implementation in clinical practice is highlighted. Specific mutations, PK and GIQ, provide relevant information for monitoring fosamprenavir-ritonavir-based HAART.


Journal of Clinical Virology | 2006

Whole blood real-time quantitative PCR for cytomegalovirus infection follow-up in transplant recipients.

Isabelle Garrigue; Sébastien Boucher; Lionel Couzi; Anne Caumont; Claire Dromer; Martine Neau-Cransac; Reza Tabrizi; Marie-Hélène Schrive; Hervé Fleury; Marie-Edith Lafon


Antiviral Therapy | 2006

Virological responses to atazanavir-ritonavir-based regimens: resistance-substitutions score and pharmacokinetic parameters (Reyaphar study).

Isabelle Pellegrin; Dominique Breilh; Jean-Marie Ragnaud; Sébastien Boucher; Didier Neau; Hervé Fleury; Marie-Hélène Schrive; Marie-Claude Saux; Jean-Luc Pellegrin; Estibaliz Lazaro; Muriel Vray


AIDS Research and Human Retroviruses | 2006

Susceptibility to Antiretroviral Drugs of CRF01_AE, CRF02_AG, and Subtype C Viruses from Untreated Patients of Africa and Asia: Comparative Genotypic and Phenotypic Data

Hervé Fleury; Thomas Toni; Nguyen Thi Hoang Lan; Pham Van Hung; Alaka Deshpande; Patricia Recordon-Pinson; Sébastien Boucher; Estibaliz Lazaro; Valérie Jauvin; Valérie Lavignolle-Aurillac; Sophie Lebel-Binay; Arnaud Chéret; Bernard Masquelier


Journal of Clinical Virology | 2005

Clonal analysis of HIV-1 variants in proviral DNA during treatment interruption in patients with multiple therapy failures

Sébastien Boucher; Patricia Recordon-Pinson; Didier Neau; Jean-Marie Ragnaud; Karine Titier; Muriel Faure; Hervé Fleury; Bernard Masquelier


Journal of Clinical Virology | 2006

Virological response to HIV-1 nucleoside/nucleotide reverse transcriptase inhibitors-based, tenofovir DF-including regimens in the ANRS Aquitaine Cohort

Eric Balestre; Michel Dupon; Sophie Capdepont; Rodolphe Thiébaut; Sébastien Boucher; Hervé Fleury; François Dabis; Bernard Masquelier


Antiviral Therapy | 2006

Antiretroviral efficacy and virological profile of a zidovudine/lamivudine/tenofovir disoproxil fumarate combination therapy in antiretroviral-naive patients.

Bernard Masquelier; Didier Neau; Sébastien Boucher; Lavignolle-Aurillac; Marie-Hélène Schrive; Patricia Recordon-Pinson; Jean-Marie Ragnaud; Hervé Fleury


AIDS Research and Human Retroviruses | 2005

Sequences of Clustered Epitopes in Gag and Nef Potentially Presented by Predominant Class I Human Leukocyte Antigen (HLA) Alleles A and B Expressed by Human Immunodeficiency Virus Type 1 (HIV-1)-Infected Patients in Vietnam

Estibaliz Lazaro; Ioannis Theodorou; Elisabeth Legrand; Patricia Recordon-Pinson; Sébastien Boucher; Corinne Capoulade; Thi Hoang Lan; Pham Van Hung; Patrice Debré; Hervé Fleury


Journal of Clinical Virology | 2006

HSV genotyping of “PCR-untyped” clinical samples

Marie-Edith Lafon; A. Gauthier; Sébastien Boucher; Muriel Faure; Isabelle Garrigue; Hervé Fleury

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Estibaliz Lazaro

Centre national de la recherche scientifique

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