乙型肝炎病毒靶序列浓度对新型肽核酸压电基因传感器阵列的影响

Effects of Hepatitis B Virus Target Concentration on New Style Peptide Nucleic Acid Piezoelectric Gene Sensor Array

目的 探索乙型肝炎病毒(HBV)靶序列浓度值对肽核酸压电基因传感器阵列杂交效应的影响。方法 压电基因传感器阵列表面先固定上1 5μmol/L的bis PNA探针,观察终浓度为1pg/L至100μg/L的HBV-DNA与探针,进行杂交所引起的频率变化及反应所需时间。并对频率下降值与靶序列浓度做线性回归,确定其线性范围。结果 靶序列浓度为1 pg/L时,传感器未能检测出任何频率的改变。随着浓度从10 pg/L增加到100μg/L,杂交反应引起的频率下降值呈先增加后趋于缓和的趋势,以10μg/L为分界线,而杂交平衡时间并没有显现出一定的变化趋势。在10pg/L 到10μg/L 的浓度范围内,浓度与频率下降值的线性回归方程为lgC= 2.7455 +0.0691×△F,相关系数r=0.9923。结论 随着靶序列浓度的升高,杂交反应引起的频率下降值呈典型饱和曲线趋势,靶序列浓度的线性检测范围为10 pg/L^10μg/L。

OBJECTIVE To investigate the effects of hepatitis B virus (HBV) target concentration on hybridization efficiency of peptide nucleic acid (PNA) piezoelectric gene sensor array. METHODS Bis-PNA probe of (1.5 μmol/L) was firstly immobilized on the surface of piezoelectric gene sensor array. Then target HBV genomic DNA varied from 1 pg/L to 100 μg/L was hybridized with the probe, and the decrease in frequency and equilibrium time of reaction were recorded and analyzed. Linear regression was done between frequency decrease and the target concentration; hence the linear range was determined. RESULTS Any frequency change could not be detected by PNA biosensor when target concentration was 1 pg/L. It enhanced firstly and then tended gently when target concentration varied from 10pg/L to 100μg/L, and 10μg/L was the watershed. And no evident change tendency was observed on equilibrium time for hybridization. The statistic linear regression equation was lgC=-2.7455+0.0691×△F among the range of target concentration from 10pg/L to 10μg/L and the correlating coefficient was 0.9923. CONCLUSIONS The frequency change caused by hybridization reaction is a typical saturation curve in accordance with the increase in target concentration. And the linear range of the target concentration is between 10 pg/L to 10μg/L.