Specific detection and differentiation of Ochrobactrum anthropi, Ochrobactrum intermedium and Brucella spp. by a multi-primer PCR that targets the recA gene

Abstract

Ochrobactrum anthropi, Ochrobactrum intermedium and Brucella spp. are phenotypically and genetically closely related pathogens that may cause disease with similar clinical presentation. Consequently, difficulties in their identification and differentiation have been reported. In this study, a sensitive recA gene-based multi-primer single-target PCR (MP-ST-PCR) was developed that allowed the specific detection and differentiation of these clinically relevant pathogens. The specificity of the assay was evaluated using a representative panel of 50 O. anthropi and 16 O. intermedium strains and the type strains of all Brucella spp. Detection limits for purified DNA from O. anthropi, O. intermedium and Brucella melitensis were 100, 10 and 100 fg, respectively. Brucella DNA was also successfully detected in various clinical specimens from a human patient with culture-proven brucellosis and from a Brucella-infected sheep and its aborted fetuses. The sensitivity of the MP-ST-PCR was comparable to that of an evaluated in-house Brucella real-time PCR assay. The developed assay closes a diagnostic gap and provides a simple but robust tool for the sensitive detection and correct identification of O. anthropi, O. intermedium and Brucella spp.

Acquired Genetic Mechanisms of a Multiresistant Bacterium Isolated from a Treatment Plant Receiving Wastewater from Antibiotic Production

“…Ochrobactrum intermedium CCUG 57381 was identified genotypically by comparative 16S rRNA and recA gene sequence analyses, since the 16S rRNA gene alone is insufficient for classification at the species level within the Ochrobactrum genus (29). The 16S rRNA gene sequence was most similar to that of the type strain of O. intermedium (accession number AM490623 [45], 99.5% nucleotide [nt] identity), and the best match for the recA sequence was also O. intermedium (AM422944, 97.7% nt identity).…”

“…The 16S rRNA genes were amplified by PCR and sequenced, targeting the hypervariable region V3 using universal primers R518 and F357, as described previously (28). The recA sequence was determined after PCR amplification using Ochr_recAF/R primers (29,30). The type strain O. intermedium LMG 3301 T , whose genome had been sequenced and the draft assembly made publically available, was selected as the reference for this study.…”

Raman Spectroscopy as a Potential Tool for Detection of Brucella spp. in Milk

“…For this reason, more sophisticated techniques, such as PCR, are in development for clinical use that can be also performed directly on complex samples such as blood or milk (1,41). However, the technique is not well established, since there is no standardization between the laboratories and sample preparation or the choice of target genes varies (60).…”

Brucella suis bacteremia misidentified as Ochrobactrum anthropi by the VITEK 2 system

“…anthropi is one of the closest Brucella relatives based on DNA, rRNA, and protein analyses [8,9]. Although both genera share phenotypic characteristics (i.e., both genera are aerobic non-fermentative, catalase and urease positive Gram-negative rods) [5], they also have meaningful differences such as their motility and susceptibility to colistin.…”