摘要
对影响寡核苷酸微阵列检测点突变的敏感性和特异性的各种因素 ,如杂交液、杂交温度、标记引物浓度及其比例等 ,进行了研究。采用不对称PCR扩增有利于敏感性提高 ;多重不对称PCR不影响杂交的特异性 ,且敏感性有所增加。对 30例肺癌标本进行寡核苷酸微阵列检测 ,发现 12例标本发生了P5 3基因点突变 ,K ras突变有 5例。与测序结果相比 ,P5 3基因突变符合率达到 80 %。由于检测样本较少且检测位点不完全 ,因而未得到K ras和P5 3基因突变与肿瘤的种类、病期及吸烟之间的明显相关性。
Different factors including hybridization solution components, hybridization temperature, and the concentration and proportion of the labelled primer, which affected the sensitivity and specificity of single mutation identification, were exploited. Asymmetric PCR increased the hybridization sensitivity, and the asymmetric multi-PCR did not affect the specificity, while the sensitivity was improved a little. Among 30 lung cancer samples detected with the oligonucleotide microarray, 12 was found P53 gene mutations and 5 had K-ras gene mutations. The P53 gene mutations identified by the oligonucleotide microarray was proved 80% same as the sequencing results. The obvious statistical relations of K-ras and P53 gene mutations with tumor type, tumor stage and smoking were not obtained because of less samples and mutation sites.
出处
《生物工程学报》
CAS
CSCD
北大核心
2002年第4期447-451,共5页
Chinese Journal of Biotechnology
基金
国家自然科学基金重点项目 (No .398890 1)
军队杰出人才基金项目资助~~
