摘要
目的 探讨变性高压液相色谱 (DHPLC)法分析肺癌患者痰标本Rb2 /p130和 p5 3突变及其作为分子诊断标记的可行性。方法 DHPLC法分析 4 7例患者痰标本 [35例肺癌 (男 2 2例 ,女13例 ,年龄 16～ 81岁 ;其中腺癌 17例 ,鳞癌 12例 ,小细胞癌 4例 ,肺泡癌 1例 ,腺鳞癌 1例 ) ,12例对照 (男 7例 ,女 5例 ,年龄 38～ 6 8岁 ;其中结核 1例 ,肺炎 1例 ,支气管扩张症 2例 ,肺泡蛋白沉积症 1例 ,结节病 1例 ,多发性软骨炎 1例 ,慢性阻塞性肺疾病 5例 ) ]Rb2 / p130基因Exon19～ 2 2和 p5 3基因Exon 5～ 9突变。结果 肺癌患者Rb2 /p130突变检出率为 2 2 86 % (8/ 35 ) ,p5 3突变检出率为2 8 5 7% (10 / 35 ) ,对照者均未检出 1例 (P =0 0 4 9,P =0 0 4 6 )。作为肺癌诊断分子标记 ,Rb2 / p130特异度 10 0 % ,灵敏度 2 2 86 % ;p5 3特异度 10 0 % ,灵敏度 2 8 5 7% ;联合检测Rb2 / p130和 p5 3特异度10 0 % ,灵敏度 5 1 4 3%。结论 联合分析痰标本Rb2 / p130和p5 3基因突变可提高诊断的灵敏度。
ObjectivesTo probe Rb2/p130 and p53 gene mutations at their hot-spots by denaturing high-performance liquid chromatography (DHPLC) analysis in sputum samplesfrom lung cancer patients and to evaluate the feasibility of these gene markers in the clinical diagnosis of lung cancer. MethodsGenomic DNAs were extracted from 47 sputum samples (35 of lung cancer,12 of benign lung disease) and their parallel peripheral blood lymphoid cells. The genomic DNAs were subjected to PCR amplification of Rb2/p130 gene at exon 19-22 and p53 gene at exon 5-9. The mutations of Rb2/p130 and p53 were detected by DHPLC by analysis of the PCR products. ResultsOf the 47 sputum samples,the Rb2/p130 gene mutation detection rates were 22.86%(8/35) in the lung cancer group and 0(0/12) in the control group ( P =0.049). The sensitivity and specificity were 22.86% and 100% respectively by using Rb2/p130 gene mutation detection as a diagnostic marker for lung cancer. p53 gene mutation detection rates were 28.57%(10/35) in the lung cancer group and 0 (0/12) in the control group ( P =0.046). The sensitivity and specificity were 28.57% and 100% respectively by using p53 gene mutation detection as a diagnostic marker for lung cancer. The sensitivity and specificity reached 51.43% and 100% respectively when Rb2/p130 and p53 gene mutations were combined as diagnostic markers for lung cancer. Conclusions Because of the low sensitivity, Rb2/p130 or p53 gene mutation detection alone is not feasible as a gene marker in the clinical diagnosis of lung cancer. The increased sensitivity,by combining the two gene markers,suggests that it may be feasible to use a panel of molecular markers such as p53,Rb2/p130,and others as diagnostic markers for lung cancer.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2004年第7期499-501,共3页
Chinese Journal of Internal Medicine
基金
国家重点基础研究发展规划项目基金资助(G19880 5 12 0 7)
