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.Brucellosis is a zoonotic disease that causes abortion, fetal death, and genital infections in animals. In humans, this highly diverse illness initially presents as fever, malaise, and myalgia and may later develop into a chronic illness affecting various organs and tissues. The causative agents of brucellosis are facultative intracellular gram-negative bacteria belonging to the genus Brucella. This genus has been further classified into six species according to host preference and pathogenicity. Genetic studies, however, indicate that the six classical species of Brucella represent a single genomospecies based upon DNA-DNA reassociation (14). In spite of this evidence, the conventional classification scheme has been maintained. Human brucellosis has been attributed to B. abortus, B. melitensis, B. suis, and B. canis and more recently to strains resembling Brucella isolated from marine mammals (13). Laboratory detection of Brucella and species identification is based largely on culture isolation and phenotypic characterization. This process is lengthy and labor-intensive and has been associated with a heightened risk of laboratory-acquired infections (7). To surmount these problems, nucleic acid amplification has been explored for the rapid detection and confirmation of Brucella.A number of nucleic acid sequences have been targeted for the development of Brucella genus-specific PCR assays, including 16S rRNA, the 16S-23S intergenic spacer region, omp2, and bcsp31 (1, 9, 11, 12). The most frequently described PCR target for the diagnosis of human brucellosis is the bcsp31 gene encoding a 31-kDa antigen conserved among Brucella spp. (8,9,15). PCR identification of Brucella strains at the species or biovar level has been more challenging. Recently, Redkar et al.(10) described real-time PCR assays for the detection of B. abortus, B. melitensis, and B. suis biovar 1. These PCR assays target the specific integration of IS711 elements within the genome of the respective Brucella species or biovar. The assays, however, were designed to be tested in separate PCRs. Using similar PCR targets, but in a multiplex format, we have developed a real-time triplex assay that permits rapid confirmation of Brucella spp., B. abortus, and B. melitensis isolates in a single test.The primers and TaqMan probes (Qiagen, Alameda, Calif

We reviewed clinical and epidemiologic features of 56 human Capnocytophaga canimorsus isolates submitted during a 32-year period to California's Microbial Diseases Laboratory for identification. An increasing number of isolates identified as C. canimorsus have been submitted since 1990. Many laboratories still have difficulty correctly identifying this species.

As crowding becomes an increasing environmental reality, humans are more frequently being exposed to zoonotic diseases as a result of encounters with "wild" animals. Domesticated animals also are exposed more frequently to wild animals; thus, increasing human exposure to once rare zoonotic illnesses. Rat-bite fever is a disease that seems to be easily recognizable by clinicians, easily identified in the clinical laboratory (if suspected), and successfully treated when the appropriate therapy is administered. Physicians should consider RBF as a possible diagnosis when fever, rash, and exposure to rats are part of the patient's history.

Burkholderia gladioli has only recently been reported to be a human pathogen. Four cases of B. gladioli infection (including bacteremia, pneumonia, and cervical adenitis) in two adults and two young children are reported. Three of these four patients were severely immunocompromised. Commercial systems were frequently unable to identify this bacterium correctly. Antimicrobial susceptibility patterns indicated that B. gladioli strains were susceptible to the quinolones, aminoglycosides, and imipenem. In vitro laboratory investigations demonstrated that B. gladioli strains were susceptible to complement-mediated lysis of pooled human serum, thus implying that healthy individuals should be immune to infection. These four cases together with three previously reported cases suggest that B. gladioli primarily causes disease in severely immunocompromised individuals. The lack of mortality associated with infection, coupled with susceptibility to serum and lack of recognizable virulence-associated factors, suggests that this species has a low pathogenic potential.