Rapid identification of Yersinia enterocolitica in blood by the 5' nuclease PCR assay.
Yersinia enterocolitica accounts for 50% of the clinical sepsis episodes caused by the transfusion of contaminated red blood cells. A 5' nuclease TaqMan PCR assay was developed to detect Y. enterocolitica in blood. Primers and a probe based on the nucleotide sequence of the 16S rRNA gene from Y. enterocolitica were designed. Whole-blood samples were spiked with various numbers of Y. enterocolitica cells, and total chromosomal DNA was extracted. When the TaqMan PCR assay was performed, as few as six bacteria spiked in 200 microliter of blood could be detected. The assay was specific and did not detect other Yersinia species. The TaqMan assay is easy to perform, takes 2 h, and has the potential for use in the rapid detection of Y. enterocolitica contamination in stored blood units.
Sen
K
K
Division of Emerging and Transfusion Transmitted Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, Maryland 20852, USA. Senk@cber.fda.gov
D004275:DNA, Ribosomal / Q000097:blood*; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D012703:Serotyping; D015009:Yersinia Infections / Q000097:blood* / Q000175:diagnosis*; D015008:Yersinia enterocolitica / Q000302:isolation & purification*
D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V