Optimization of real-time PCR assay for rapid and sensitive detection of eubacterial 16S ribosomal DNA in platelet concentrates.
A real-time PCR assay was developed for rapid detection of eubacterial 16S ribosomal DNA in platelet concentrates. The sensitivity of this assay can be hampered by contaminating DNA in the PCR reagents. Digestion of the PCR reagents with Sau3AI prior to PCR amplification was effective in eliminating this contaminating DNA without affecting the sensitivity of the assay.
Mohammadi
Tamimount
T
Sanquin Blood Bank North West Region, Amsterdam, The Netherlands.
Reesink
Henk W
HW
Vandenbroucke-Grauls
Christina M J E
CM
Savelkoul
Paul H M
PH
D001792:Blood Platelets / Q000382:microbiology*; D004269:DNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000032:analysis* / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000378:metabolism; D013997:Time Factors
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D019914:Taq Polymerase; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases