摘要
为解决常规ＤＮＡ单链构象多态性分析（ＳＳＣＰ）使用放射性同位素及操作繁杂等问题，应用溴化乙锭（ＥＢ）染色的非同位素检测方法对１６例卵巢癌及５例正常胎盘组织中ｐ５３基因的第５、６外显子（Ｅｘｏｎ５、６）的聚合酶链反应（ＰＣＲ）扩增产物进行了分析，并结合ＤＮＡ测序技术进行验证。正常组织未发现异常，１６例卵巢癌ＳＳＣＰ分析显示３例异常，ＤＮＡ测序证实其中２例为核苷酸突变，１例为核苷酸插入。采用ＥＢ染色的非同位素ＰＣＲ－ＳＳＣＰ，方法简便、省时、准确，具有良好的实用性和广泛的使用价值。
or solving the problems of using radioactive isotope and operational complexity in routine single-strand conformation polymorphism analysis ( SSCP ),a new technique using ethidium bromide ( EB)staining nonisotopic SSCP and polymerase chain reaction(PCR)was developed.The exon 5 and 6 of p53 gene in 16 cases of ovarian cancer and 5 cases of normal placenta were examined with this technic. The results were confirmed by DNA direct sequencing. No alterations were detected in normal placen-tas.3/16 of the ovarian cancer cases showed abnormal in PCR-SSCP. The DNA sequencing data con-firmed mutations of DNA in 2 cases and insertion of nucleotied‘G’in another case. The results suggest-ed that the EB staining nonisotopic PCR-SSCP is a simple,time saving and accurate method which may be used in routine PCR-SSCP.
