Real-time PCR for detection of Brucella spp. DNA in human serum samples.
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.
Debeaumont
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Laboratoire de Bactériologie, Centre Hospitalier Universitaire, Faculté de Médecine, Université Joseph Fourier, BP 217, 38043, Grenoble cedex 9, France.
Falconnet
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Maurin
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D000328:Adult; D000368:Aged; D016470:Bacteremia / Q000175:diagnosis / Q000382:microbiology*; D001426:Bacterial Proteins / Q000235:genetics; D002002:Brucella / Q000235:genetics / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis / Q000382:microbiology*; D002648:Child; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000097:blood*; D005260:Female; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping / Q000379:methods
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D008565:Membrane Proteins