Abstract：ObjectiveTo develop a modified molecular beacon real-time PCR assay based on Homo-Tag Assisted Non-Dimer (HAND) system for detection of Laribacter hongkongensis, and optimize and evaluate the assay. MethodsBased on the consensus sequences of 16SrDNA gene published on GeneBank, primers and molecular beacon probe of Laribacter hongkongensis were designed, the homologous tail sequences were added to the 5’ end to prevent or reduce the formation of primer dimer．The fluorescence PCR system was constructed by optimizing the PCR parameters like concentrations of homologous tail primers and specific primers as well as reaction temperature. ResultsThe modified molecular beacon real-time PCR assay based on HAND system was developed, in which the primer dimer was significantly reduced. The sensitivity of the assay was 20 CUF/ml. ConclusionsThe real-time PCR assay based on HAND system has the advantage of rapidity, sensitivity and specificity. It can be applied to the rapid detection of Laribacter hongkongensis.
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2015, Vol. 22 
