应用压电生物传感器检测HBV感染相关SNP位点的实验研究

SNP Detection Relative to HBV Infection by DNA Piezoelectric Biosensor

目的建立压电生物传感器单核苷酸多态性(SNP)检测方法,检测乙型肝炎病毒(HBV)感染相关SNP位点ESR1 T29C,期望为临床提供一种准确、可靠、快速、简便的SNP检测方法。方法以合成DNA序列target-A和target-G为检测靶序列,建立压电传感器检测SNP的模式实验,并确定最低检测灵敏度;收集临床患者外周血样本,提取基因组DNA,经测序方法鉴定SNP位点基因型,对不同基因型样本ESR1 T29C位点所在片段进行PCR扩增后,应用压电生物传感器进行检测。结果检测靶序列为target-A时,频率下降(416.0±21.5)Hz;检测靶序列为target-G时,频率下降(9.4±5.0)Hz,两者频率下降值差异有统计学意义(P<0.01),并最低可检测到2×10-11mol/L,以基因组DNA PCR扩增产物为检测靶序列时,基因型TT频率下降(109.4±13.4)Hz;TC型频率下降(52.0±11.4)Hz;CC型频率下降(7.2±4.5)Hz,3者频率下降值差异有统计学意义(P<0.05)。结论该方法能够特异、灵敏地检出临床标本中SNP位点的不同基因型。

OBJECTIVE To establish a SNP detection method by DNA piezoelectric biosensor and detect a SNP relative to HBV infection. METHODS To establish a model experiment with synthesis DNA sequences as target and find the lowest sensitivity. After extraction of genome DNA from inpatient blood sample, the SNP sites located in ESR1 gene region in samples were detected by SNP detecting method established. RESULTS The frequency shift of target-A was 416, 0±21.5Hz, the frequency shift of target-G was 9.4±5.0Hz. And it could be detected that the lowest sensitivity of target-A was 2×10^-11 mol/L. The three genotypes of blood samples, TT, TC and CC, had different frequency shifts, 109.4±13.4Hz, 52.0±11.4Hz and 7. 2±4.5Hz, respectively. CONCLUSIONS SNP in blood sample could be detected specifically and sensitively by DNA piezoelectric biosensor.