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本文以野生型的乙型肝炎病毒(HBV)核酸片段为研究对象,利用无标记的分子信标及核酸染料SYBR Green I,建立了一种高灵敏、高选择性的特定序列核酸检测方法.在优化条件下,目标DNA浓度为4×10-11~400×10-11 mol/L之间时,SYBR Green I的荧光强度(ΔI)与目标DNA的浓度(C)具有良好的线性关系,其拟合的回归方程为ΔI=1.9556 C+31.4659(R2=0.9956),方法检测限(3ζ)为2×10-11 mol/L.该方法操作简单、检测速度快、灵敏度高、重现性好、检出限低.利用该方法,结合不对称PCR技术,实现了对HBV的定量检测.
In this paper, wild-type hepatitis B virus (HBV) nucleic acid fragments as the research object, the use of unlabeled molecular beacon and nucleic acid dye SYBR Green I, established a highly sensitive and highly selective sequence-specific nucleic acid detection method. Under optimal conditions, the fluorescence intensity (ΔI) of SYBR Green I and the concentration of target DNA (C) had a good linear relationship when the target DNA concentration was between 4 × 10-11 ~ 400 × 10-11 mol / L. The regression equation of fitting was ΔI = 1.9556 C + 31.4659 (R2 = 0.9956), the limit of detection (3ζ) was 2 × 10-11 mol / L. The method was simple, rapid, sensitive and reproducible , The detection limit is low.This method, combined with asymmetric PCR technology, to achieve a quantitative detection of HBV.