Comparison of three different PCR methods for detection of Brucella spp in human blood samples.
In the diagnosis of human brucellosis, PCR could be a more sensitive technique than blood cultures and more specific than conventional serological tests. We compared three different PCR methods for the detection of Brucella spp. and we studied whether human genomic DNA affect the sensitivity of three primer pairs for the detection of Brucella DNA in a peripheral-blood PCR assay. These three pairs of primers amplified three different fragments included in: (i). a gene encoding a 31-kDa Brucella abortus antigen (primers B4/B5), (ii). a sequence 16S rRNA of B. abortus (primers F4/R2), and (iii). a gene encoding an outer membrane protein (omp-2) (primers JPF/JPR). The three primers assayed showed a difference in sensitivity for detecting purified Brucella DNA, ranging between 8 fg and 20 pg. However, the sensitivity of the primers F4/R2 and B4/B5 was affected by the presence of human DNA while the primers JPF/JPR were not. Therefore, although the sensitivity of PCR using primers F4/R2 is affected by human DNA, they are still the most sensitive and they could provide a useful tool for the diagnosis of human brucellosis.
Navarro
E
E
Experimental Research Unit, Albacete General Hospital, c/Hermanos Falcó s/n, 02006, Albacete, Spain.
Escribano
J
J
Fernández
J
J
Solera
J
J
D002002:Brucella / Q000235:genetics / Q000302:isolation & purification*; D017347:Brucella melitensis / Q000235:genetics / Q000302:isolation & purification; D002006:Brucellosis / Q000175:diagnosis*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000097:blood*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial