ABSTRACT
OBJECTIVE To develop a new rapid method of real time fluorescence quantitative Polymerase chain reaction(PCR) to detect Polyoma virus BK(BKV) with high sensitivity,specificity,stability and lower price using the self-quenched probe.METHODS DNA segment of BKV was cloned into the vector pGEM-11Zf.The recombinant vector pGEM-11Zf-BK was used as standard template.The self quenched probe was designed according to the cloned gene sequence.The PCR reaction system was optimized and the method established was evaluated.RESULTS Restriction maps showed that the structure of recombinant plasmid pGEM-11Zf-BK was the same as expected.Methodology analysis suggested that self-quenched probe fluorescent quantitative PCR detective method of BKV be successfully established with high sensitivity,specificity and accuracy.CONCLUSIONS The self-quenched probe fluorescence quantitative PCR detective method for BKV has been developed successfully with high sensitivity,specificity and accuracy.