Comparison of seven commercial DNA extraction kits for the recovery of Brucella DNA from spiked human serum samples using real-time PCR.
We compared the relative recovery of extraction of bacterial DNA from serum using seven commercial kits (UltraClean DNA BloodSpin Kit, Puregene DNA Purification System, Wizard Genomic DNA Purification Kit, High Pure PCR Template Preparation Kit, GFX Genomic Blood DNA Purification Kit, NucleoSpin Tissue Kit, and QIAamp DNA Blood Mini Kit). Human serum samples were spiked with known concentrations of Brucella melitensis Rev 1; the DNA was extracted and tested in genus-specific LightCycler polymerase chain reaction (PCR). The UltraClean DNA BloodSpin Kit proved to be as sensitive as the QIAamp DNA Blood Mini Kit isolation method and could detect down to 100 fg of DNA, though only the former had no contamination. All the other procedures yielded DNA isolation results that were less sensitive and were always contaminated. Our results show that the UltraClean DNA Blood Spin Kit was the commercially available assay tested that yielded the best sensitivity, purity, and lack of contamination for Brucella DNA isolation from serum.
Queipo-Ortuño
M I
MI
Biochemistry and Molecular Biology Department, Faculty of Medicine, Universidad de Málaga, Campus de Teatinos s/n, 29071 Málaga, Spain. iqueipo@uma.es
Tena
F
F
Colmenero
J D
JD
Morata
P
P
D001431:Bacteriological Techniques / Q000379:methods*; D002002:Brucella / Q000235:genetics / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D044967:Serum / Q000382:microbiology*
D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic