Study on the development of a real-time quantitative polymerase chain reaction assay to detect Rickettsia / 中华流行病学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To develop a real-time quantitative polymerase chain reaction(PCR) assay for detecting Rickettsia rickettsii.</p><p><b>METHODS</b>The primers and TaqMan-MGB probe were designed according to the ompB gene of R. rickettsii. A DNA fragment of ompB gene amplified from R. rickettsii by PCR was used as a standard template for the development of the method.</p><p><b>RESULTS</b>5 copies of ompB fragments of R. rickettsii were detected. The genomic DNA of R. rickettsii was detected by the developed quantitative PCR assay. However, the genomic DNA from another rickettsial or bacterial agent was not determined. Through this developed method, the positive results were obtained from the animals and cells, artificially infected with R. rickettsii.</p><p><b>CONCLUSION</b>The real-time quantitative PCR assay seemed to be highly sensitive and specific which might be used to rapidly detect R. rickettsia DNA in various samples and to early diagnose patients infected by R. rickettsii.</p>