乙肝病毒突变位点的寡核苷酸与肽核酸阵列杂交检测

目的：比较利用寡核苷酸阵列和肽核酸阵列检测乙肝病毒突变位点。方法：针对乙肝病毒常见的1762A-T与1764G-A联合突变、1858C-T、 1896G-A 3个突变位点，设计寡脱氧核苷酸和肽核酸检测探针，探针经固定后分别与不对称PCR荧光标记的靶DNA杂交，扫描分析荧光信号强度，DNA序列分析3位点的变异情况。结果：对于寡核苷酸芯片，1762与1764联合位点、1858以及1896位点的野生型探针与突变探针荧光强度的比值分别为1.44、1/1.24、1.05。对于肽核酸芯片，相应位点的野生型探针与突变探针荧光强度的比值分别为2.8、1/3.02、1.71。DNA序列分析结果表明该样本中1858位点为突变位点，1896以及1762与1764联合位点不存在突变。结论：与寡脱氧核苷酸探针相比，肽核酸与靶DNA杂交结合能力和识别单碱基错配能力均高于相应的寡脱氧核苷酸，二者杂交结果与测序结果一致。

Hybridization detection of mutation sites in hepatitis B virus using oligodeoxynucleotide and PNA array

Objective: To compare the oligonucleotide array and PNA array in determining mutation of HBV.Methods: Oligonucleotide and PNA probes which targeted 1762A-T and 1764G-A,1858C-T and 1896G-A mutations and wild sites of HBV were designed and immobilized on aldehyde modified glass slides after synthesis.Target DNA was obtained by asymmetric PCR with a tamara-labeled upper primer.Result was produced by scanning and quantifying analysis after hybridization with labeled target DNA.Meanwhile,the PCR product was sequenced to analyze three mutation sites.Results: The ratio of fluorescence signals for 17621764 double site, 1858 and 1896 sites(wild type: mutation type) was 1.44,1/1.24 and 1.05 in the oligonucleotide array and 2.8,1/3.02,1/1.71 in the PNA array.1858T mutation and 1762A-1764G,1896G wild sites were found in DNA sequencing.Conclusions: The oligonucleotide array and the PNA array could successfully detect the three mutation sites of HBV,which is in accordance with the result of DNA sequencing.PNA shows a high priority in attaching the target DNA and identifying the single-base mismatch.