Detection of Mycobacterium tuberculosis by Fluorescence Chip of Molecular Beacon Probe / 中华医院感染学杂志

ABSTRACT

OBJECTIVE To have a research on the detection of Mycobacterium tuberculosis by fluorescence chip of molecular beacon probe.METHODS Fluorescence chip for the detection of M.tuberculosis was designed,based on the better bonding force and specificity of molecular beacon and target molecule.Hybridization efficiency was optimized,which included the concentration and purity of molecular beacon probe,hybridization temperature,hybridization time,the prescription of hybridization solution and the comparing of molecular beacon fluorescence signals of 4 molecules.RESULTS Molecular beacon fluorescence signal approached the highest when the final concentration of Mg was 5 mmol/L and the amplification efficiency of PCR approached the highest.Half extended molecular beacon probe could bind nice with chip when it was middle labeled with quench group.The results of 3 hybridization solutions were as follows 0.15% SDS better than 0.2% SDS,10?SSC better than 5?SSC.Strong signal of hybridization reaction could be detected when the concentration of Cy3-BDH molecular beacon probe was 15 ?mol/L,the concentration of FAM-DABCLYE probe was 9 ?mol/L and the reaction time was 7 h.The efficiency of fluorescence quenching was higher when the molecule was consisted of Cy3-BDH,other than Cy3-DABCLYE.CONCLUSIONS The design of half extended molecular beacon probe tactfully makes use of the advantages of the high bonding force and specificity of molecular beacon probe on the chip technology.Fluorescence chip of molecular beacon for M.tuberculosis has satisfactory detecting rate,specificity and sensibility.