Specific detection and differentiation of Ochrobactrum anthropi, Ochrobactrum intermedium and Brucella spp. by a multi-primer PCR that targets the recA gene.
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.
Scholz
Holger Christian
HC
Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, D-80937 Munich, Germany.
Pfeffer
Martin
M
Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, D-80937 Munich, Germany.
Witte
Angela
A
Institute of Microbiology and Genetics, University of Vienna, Dr Bohr-Gasse 9, A-1030 Vienna, Austria.
Neubauer
Heinrich
H
Friedrich Loeffler Institute, Institute of Bacterial Infections and Zoonoses, Federal Research Institute for Animal Health, Naumburger Strasse 96a, D-07743 Jena, Germany.
Al Dahouk
Sascha
S
RWTH University of Aachen, Department of Internal Medicine III, Pauwelsstraße 30, D-52074 Aachen, Germany.
Wernery
Ulrich
U
Central Veterinary Research Laboratory, PO Box 597, Dubai, United Arab Emirates.
Tomaso
Herbert
H
Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, D-80937 Munich, Germany.
D000818:Animals; D015373:Bacterial Typing Techniques; D002002:Brucella / Q000235:genetics / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000382:microbiology*; D006801:Humans; D042041:Ochrobactrum / Q000235:genetics / Q000302:isolation & purification*; D020621:Ochrobactrum anthropi / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011938:Rec A Recombinases / Q000235:genetics*; D012680:Sensitivity and Specificity; D012756:Sheep
D017931:DNA Primers; D004269:DNA, Bacterial; D011938:Rec A Recombinases