Real-Time Multiplex PCR Assay for Detection of Brucella spp., B. abortus, and B. melitensis

ABSTRACT The identification of Brucella can be a time-consuming and labor-intensive process that places personnel at risk for laboratory-acquired infection. Here, we describe a real-time PCR assay for confirmation of presumptive Brucella isolates. The assay was designed in a multiplex format that will allow the rapid identification of Brucella spp., B. abortus, and B. melitensis in a single test.

[1]  M. Queipo-Ortuño,et al.  Development and Evaluation of a PCR-Enzyme-Linked Immunosorbent Assay for Diagnosis of Human Brucellosis , 2003, Journal of Clinical Microbiology.

[2]  T. Carpenter,et al.  Time-Space Clustering of Human Brucellosis, , 2002 .

[3]  B. Drasar,et al.  Detection of Brucella melitensis and Brucella abortus by DNA amplification. , 1992, The Journal of tropical medicine and hygiene.

[4]  J. Solera,et al.  Comparison of three different PCR methods for detection of Brucella spp in human blood samples. , 2002, FEMS immunology and medical microbiology.

[5]  K. Bourantas,et al.  Serum Is the Preferred Clinical Specimen for Diagnosis of Human Brucellosis by PCR , 2001, Journal of Clinical Microbiology.

[6]  T. Carpenter,et al.  Time-Space Clustering of Human Brucellosis, California, 1973–1992 , 2002, Emerging infectious diseases.

[7]  R. Borrow,et al.  Contamination and Sensitivity Issues with a Real-Time Universal 16S rRNA PCR , 2000, Journal of Clinical Microbiology.

[8]  C. Gamazo,et al.  Specific detection of Brucella DNA by PCR , 1995, Journal of clinical microbiology.

[9]  C. D. Miller,et al.  A twenty-five year review of laboratory-acquired human infections at the National Animal Disease Center. , 1987, American Industrial Hygiene Association journal.

[10]  J. Derr,et al.  Species-specific sequences at the omp2 locus of Brucella type strains. , 1996, International journal of systematic bacteriology.

[11]  L. Herman,et al.  Direct detection of Brucella spp. in raw milk by PCR and reverse hybridization with 16S-23S rRNA spacer probes , 1996, Applied and environmental microbiology.

[12]  S. Rose,et al.  Real-time detection of Brucella abortus, Brucella melitensis and Brucella suis. , 2001, Molecular and cellular probes.

[13]  A. Bollen,et al.  Human Neurobrucellosis with Intracerebral Granuloma Caused by a Marine Mammal Brucella spp. , 2003, Emerging infectious diseases.

[14]  A. Cloeckaert,et al.  Nucleotide sequence and expression of the gene encoding the major 25-kilodalton outer membrane protein of Brucella ovis: Evidence for antigenic shift, compared with other Brucella species, due to a deletion in the gene , 1996, Infection and immunity.

[15]  F. Grimont,et al.  Brucella, a monospecific genus as shown by deoxyribonucleic acid hybridization , 1985 .

[16]  M. Queipo-Ortuño,et al.  Specificity of a Polymerase Chain Reaction Assay of a Target Sequence on the 31-Kilodalton Brucella Antigen DNA Used to Diagnose Human Brucellosis , 2001, European Journal of Clinical Microbiology and Infectious Diseases.