Detection and differentiation of the six Brucella species by polymerase chain reaction.
BACKGROUND
Brucelosis is a severe acute febrile disease caused by bacteria of the genus Brucella. Its current diagnosis is based on clinical observations that may be complemented by serology and microbiological culture tests; however, the former is limited in sensitivity and specificity, the latter is time consuming. To improve brucelosis diagnosis we developed a test which is specific and sensitive and is capable of differentiating the six species of Brucella.


METHODS
Four primers were designed from B. abortus sequences at the well-conserved Omp2 locus that are able to amplify the DNAs of all six species of Brucella.


RESULTS
Our test detected all six species of Brucella. Their differentiation resulted directly from differences in the amplification patterns or was achieved indirectly using a RFLP present in one of the PCR products. The sensitivity and specificity of the new test were then determined; it was applied successfully in confirming the diagnosis of a patient whose clinical history and serology indicated infection with Brucella.


CONCLUSIONS
The results make possible the use of a PCR test for Brucella detection and differentiation without relying on the measurement of the antibodies or microorganism culture. Our first results showed that the PCR test can confirm the presence of Brucella in blood samples of infected patients.
Sifuentes-Rincón
A M
AM
Departamento de Bioquímica, Facultad de Medicina, Universidad Autónoma de Nuevo León, Monterrey, México.
Revol
A
A
Barrera-Saldaña
H A
HA
D002002:Brucella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D002003:Brucella abortus / Q000235:genetics / Q000302:isolation & purification; D017347:Brucella melitensis / Q000235:genetics / Q000302:isolation & purification; D002006:Brucellosis / Q000097:blood / Q000175:diagnosis*; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial