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随着分子生物学的迅速发展,使肿瘤的病因、诊断及治疗等领域进入了基因水平。癌是基因性疾病,各种致癌因素诱发癌基因激活和抗癌基因失活及产物异常是癌变的首要因素,这些变化的基础就是基因异常改变。传统的Southern印迹杂交法已无法满足对癌细胞的基因分析,特别是对基因异变的检测。PCR直接测序分析是在PCR扩增基因组DNA序列的基础上直接进行基因核苷酸序列分析的方法。它能检测肿瘤细胞单个核苷酸的改变,且省去了基因克隆等繁琐步骤,是检测基因突变最灵敏、最简单、最快速和最精确的方法。PCR直接测序已广泛用于癌基因和抗癌基因突变的研究、遗传鉴定、传染性疾病的诊断。核苷酸序列的测定似乎可以说是分子生物学最重要的技术,PCR直接测序方法的不断完善,无疑对分子肿瘤学的研究,提供更多、更准确的信息。
With the rapid development of molecular biology, the etiology, diagnosis and treatment of tumors have entered the genetic level. Cancer is a genetic disease. The oncogenic activation and anti-oncogene inactivation and abnormal production of various cancer-causing factors are the primary factors of canceration. The basis of these changes is genetic abnormalities. Traditional Southern blot hybridization can no longer meet the genetic analysis of cancer cells, especially the detection of genetic mutations. PCR direct sequencing analysis is a method of directly performing nucleotide sequence analysis of genes based on PCR amplification of genomic DNA sequences. It can detect changes in individual nucleotides of tumor cells, and eliminates the cumbersome steps of gene cloning. It is the most sensitive, simple, fastest, and most accurate method for detecting gene mutations. Direct PCR sequencing has been widely used for the study of oncogenes and anti-oncogene mutations, genetic identification, and diagnosis of infectious diseases. The determination of nucleotide sequences seems to be the most important technique in molecular biology. The continuous improvement of PCR direct sequencing methods undoubtedly provides more and more accurate information on molecular oncology research.