[Detection of bacterial DNA in blood with three pairs of primers].
OBJECTIVE
To develop a stable and sensitive method to detect gut-derived bacterial DNA present in the blood.


METHODS
Blood culture and PCR were performed simultaneously when temperature of patients (n = 23) was higher than 38.5 degrees C after laparotomy. PCR was performed after DNA extraction, target genes being beta-lactosidase gene of Escherichia coli(E. coli), glutamine synthase gene of Bacteroides Fragilitas, and 16SrRNA gene of most pathogenic bacteria. DNA of standard strains of E. coli and Bacteroides Fragilitas served as positive controls, blank control as negative controls, and healthy volunteers (n = 20) served as normal controls. We performed PCR twice with part of the samples (n = 20) in order to test the repetition of this method.


RESULTS
The repetition rate of PCR was 95%. Three of blood cultures were positive (13.0%, 3/23), the corresponding positive ratio of PCR was 43.5% (10/23) which was higher than that of the blood culture (P = 0.016).


CONCLUSIONS
PCR is more sensitive than blood culture in detection of bacteremia.
Wen
L
L
Department of Surgery, PUMC Hospital, CAMS and PUMC, Beijing 100730, China.
Tang
W
W
Jiang
Z
Z
Wang
S
S
Liang
C
C
Zhang
Y
Y
D018988:Bacterial Translocation; D017931:DNA Primers; D004269:DNA, Bacterial / Q000097:blood*; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D005974:Glutamate-Ammonia Ligase / Q000235:genetics; D006026:Glycoside Hydrolases / Q000235:genetics; D006403:Hematologic Tests; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D006026:Glycoside Hydrolases; D005974:Glutamate-Ammonia Ligase