核酸杂交的二次酶放大镧系元素发光时间分辨荧光分析
Two-round enzyme-amplified lanthanide luminescence time-resolved fluorescence assay for nucleic acid hybridization
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摘要: 目的 建立一种超灵敏的用于核酸杂交分析的方法。方法 通过二次酶放大、PCR扩增、Tb螯合物和时间分辨测量技术,测定前列腺特异抗原(PSA)DNA。结果 靶PSADNA测定的标准曲线线性范围大于两个数量级;灵敏度为10pmol/L(0.5fmol/孔);当靶PSADNA为10、30和60pmol/L时,准确度和精密度分别为77%~95%和12.9%~15.3%。结论 本法是一种超灵敏的、简捷和快速的全新分析方法,有很好的应用前景。Abstract: Objective To investigate a supersensitive for nucleic acid hybridization assay. Methods Using the two-roung enzyme-amplified action, PCR amplification, Tb chelate and time-resolved detection technology, target PSA DNA was detected. Results The range of the standard curve for detecting target PSA DNA is more than two order of magnitude; the sensitivity was 10pmol/L (0.5fmol/well); the accuracy and the precision was 77%~95% and 12.9%~15.3% respectively when the target PSA DNA was 10,30 and 60pmol/L. Conclusion The method is a supersensitive and new assay technology which has a good prospect of applications.
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