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2 results on '"Pierre-Alain Falconnet"'

1. Real-time PCR for detection of Brucella spp. DNA in human serum samples

Author

Pierre-Alain Falconnet, M Maurin, C Debeaumont, Laboratoire de Bactériologie, CHU Grenoble, Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), and Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)

Subjects
Male, Bacteremia, Brucellaceae, Polymerase Chain Reaction, law.invention, Serology, law, MESH: Child, Child, MESH: Bacteremia, Yersinia enterocolitica, MESH: Bacterial Proteins, Polymerase chain reaction, MESH: Aged, 0303 health sciences, MESH: Middle Aged, biology, MESH: Brucella, General Medicine, Middle Aged, Infectious Diseases, Real-time polymerase chain reaction, Female, MESH: Membrane Proteins, Adult, DNA, Bacterial, MESH: DNA Primers, Microbiology (medical), Brucella, Brucellosis, Microbiology, 03 medical and health sciences, Bacterial Proteins, MESH: Brucellosis, medicine, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Serotyping, Aged, DNA Primers, 030304 developmental biology, MESH: Humans, 030306 microbiology, Membrane Proteins, MESH: Serotyping, MESH: Adult, MESH: Polymerase Chain Reaction, biology.organism_classification, medicine.disease, MESH: DNA, Bacterial, Virology, MESH: Male, genomic DNA, MESH: Female
Abstract

Presented here are the results of an evaluation of an in-house real-time PCR assay for the rapid and specific diagnosis of human brucellosis. The assay was based on direct amplification from serum samples of a 169-bp portion of bcsp31, a gene found in all Brucella species and biovars. Species specificity and selectivity of this real-time PCR assay were evaluated using genomic DNA from 15 Brucella strains and 42 non-Brucella strains, and the results were 100%. Among 17 culture-proven brucellosis patients, sera from 11 gave a positive amplification signal, corresponding to a sensitivity of 64.7%. In contrast, negative results were obtained for all sera from 60 control patients, corresponding to a specificity of 100%. The results indicate this test is well adapted for definite confirmation of brucellosis cases, when Brucella cultures remain sterile and serological tests demonstrate the presence of cross-reacting antibodies against Brucella sp. and Yersinia enterocolitica O:9 antigens.

Published
2005
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2. Quantitative Real-Time Legionella PCR for Environmental Water Samples: Data Interpretation

Author

M. Reyrolle, François Vandenesch, Nicole Weill, Jerome Etienne, Janine André, Pierre-Alain Falconnet, Sophie Jarraud, Max Maurin, Philippe Joly, Centre National de Référence des Légionella, Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Bactériologie, CHU Grenoble, CARSO-Laboratoire Santé Environnement Hygiène de Lyon (CARSO-LSEHL), CARSO-LSEHL, Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), and Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)

Subjects
Genus Legionella, Colony Count, Microbial, Fresh Water, Public Health Microbiology, 010501 environmental sciences, 01 natural sciences, Applied Microbiology and Biotechnology, Legionella pneumophila, Polymerase Chain Reaction, law.invention, law, RNA, Ribosomal, 16S, MESH: Water Supply, Polymerase chain reaction, 0303 health sciences, Ecology, biology, MESH: Air Conditioning, Reference Standards, MESH: RNA, Ribosomal, 16S, MESH: Fresh Water, MESH: Laboratories, MESH: Reference Standards, Biotechnology, DNA, Bacterial, Legionella, Sensitivity and Specificity, Microbiology, 03 medical and health sciences, Environmental water, Water Supply, Air Conditioning, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Letters to the Editor, MESH: Colony Count, Microbial, 0105 earth and related environmental sciences, Chromatography, 030306 microbiology, Data interpretation, MESH: Polymerase Chain Reaction, Ribosomal RNA, 16S ribosomal RNA, biology.organism_classification, MESH: DNA, Bacterial, MESH: Legionella pneumophila, MESH: Sensitivity and Specificity, Culture Media, respiratory tract diseases, MESH: Culture Media, Laboratories, Food Science, MESH: Legionella
Abstract

Quantitative Legionella PCRs targeting the 16S rRNA gene (specific for the genus Legionella ) and the mip gene (specific for the species Legionella pneumophila ) were applied to a total of 223 hot water system samples (131 in one laboratory and 92 in another laboratory) and 37 cooling tower samples (all in the same laboratory). The PCR results were compared with those of conventional culture. 16S rRNA gene PCR results were nonquantifiable for 2.8% of cooling tower samples and up to 39.1% of hot water system samples, and this was highly predictive of Legionella CFU counts below 250/liter. PCR cutoff values for identifying hot water system samples containing >10 3 CFU/liter legionellae were determined separately in each laboratory. The cutoffs differed widely between the laboratories and had sensitivities from 87.7 to 92.9% and specificities from 77.3 to 96.5%. The best specificity was obtained with mip PCR. PCR cutoffs could not be determined for cooling tower samples, as the results were highly variable and often high for culture-negative samples. Thus, quantitative Legionella PCR appears to be applicable to samples from hot water systems, but the positivity cutoff has to be determined in each laboratory.

Published
2006
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