Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 9/15/2003
Publication Date: N/A
Technical Abstract: There has recently been increased interest in the development of molecular-based diagnostic techniques for the identification and fingerprinting of Brucella isolates. One impediment for this technology is sample preparation, since most methods (especially PCR-based methods) are very sensitive to inhibitors from blood, milk, tissues, or foods. Culturing the bacteria is an effective way to eliminate inhibitors, but it is time-consuming. Alternatively, DNA extraction and purification from the inhibitors is also time consuming and may lead to cross-contamination of samples. We describe the use of commercially available FTA cards as a rapid method for preparing Brucella DNA for PCR and other enzymatic reactions. These cards, commonly used for forensic analysis of blood and buccal samples, consist of filter paper impregnated with solubilizers and detergents. When a sample is spotted on the card the cells or bacteria are lysed and the DNA binds to the filter. Contaminants and inhibitors are easily removed by a few wash steps. We demonstrated that the treated samples are readily amplified by PCR without any significant loss of sensitivity.