Establishment and application of real-time fluorescence polymerase chain reaction based on the TaqMan probes for detection of Yersinia pestis / 中华流行病学杂志

ABSTRACT

<p><b>OBJECTIVE</b>Establishing and developing methods for quick, special and of Yersinia pestis (Y. pestis).</p><p><b>METHODS</b>Using real-time fluorescence polymerase chain reaction (PCR) based on TaqMan technology with UDG anti-contamination systems and ROX reference dye, we established a method to detect Y. pestis. Probes and primers from pla, caf1, hms and inv gene were designed. An internal positive control was added to the reaction mixture in order to detect the presence of PCR inhibitors that were often found in biological samples. Sensitivity was evaluated by serial dilution of colons, internal template and Y. pestis strains while specificity was confirmed by amplifying real DNAs from bacteria, the representative Y. pestis strains and blind assay.</p><p><b>RESULTS</b>The methods used could provide quick, special and sensitive detection of Y. pestis, with sensitivities of pla, f1, hms as 10, 1 and 1 copie(s) respectively.</p><p><b>CONCLUSION</b>The newly developed technologies seemed to be well suited to identify the Y. pestis in case of emergency, bio-terrorist attack and surveillance of the epidemics.</p>