Development and evaluation of a rapid DNA preparation method for PCR-based DNA virus detection

Abstract

We describe a simple, rapid and resource-saving method of DNA preparation from cultured cells, sera and animal tissues for PCR-based DNA virus detection. The method does not require the proteinase K, ethanol or phenol/chloroform used in conventional methods, and the entire procedure is performed in the same tube, reducing possible cross contamination between samples and the expense of laboratory ware. The protocol utilizes guanidine HCl and sodium dodecyl sulfate successively to lyse cells and dissociate proteins from nucleic acid at high temperature, and precipitates SDS and proteins at low temperature while reducing guanidine HCl concentration sufficiently to permit PCR-based virus detection. This method is extremely low cost, high sensitivity and provides a quick and effective way for clinical and laboratory virus detection, and is especially useful for simultaneous analysis of a large number of samples.