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目的:验证咽拭子法检测EB病毒(EBV)潜伏膜蛋白1(LMP1)在鼻咽癌诊断中应用的可行性;检测LMP1野生型及30bp缺失变异型在鼻咽癌中的分布情况。方法:用咽拭子法收集鼻咽癌组及对照组鼻咽部脱落细胞,微量提取DNA,经PCR扩增人β珠蛋白基因序列验证咽拭子法的可行性,用特异性引物扩增出特异的LMP1序列,验证其在鼻咽癌诊断中的意义。测序分析LMP1变异情况。结果:咽拭子合格率为96.4%,LMP1基因作为鼻咽癌的检测指标,灵敏度为91.7%,特异性为95.6%,其中变异型LMP1在鼻咽癌中的表达频率为80.6%,野生型为11.1%。结论:咽拭子法可作为一种基因检测手段,LMP1基因的检测可以协同EB病毒壳抗原(EBVCA)IgA作为鼻咽癌诊断的辅助指标,在LMP1(+)的鼻咽癌患者中LMP1-30bp缺失突变普遍存在。
Objective: To verify the feasibility of using throat swab method to detect Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) in the diagnosis of nasopharyngeal carcinoma and to detect the distribution of LMP1 wild type and 30bp mutant in nasopharyngeal carcinoma. Methods: Throat swabs were used to collect nasopharyngeal exfoliated cells from nasopharyngeal carcinoma group and control group. DNA was extracted by micro-extraction. The feasibility of throat swab method was verified by PCR amplification of human β-globin gene sequence. A special LMP1 sequence to verify its significance in the diagnosis of nasopharyngeal carcinoma. Sequencing analysis of LMP1 mutation. Results: The pass rate of pharyngeal swab was 96.4%. The detection rate of LMP1 gene was 91.7% and the specificity was 95.6%. The frequency of LMP1 gene expression in NPC was 80.6% 11.1%. Conclusion: The throat swab method can be used as a genetic test method. The detection of LMP1 gene can cooperate with Epstein-Barr virus capsid antigen (EBVCA) IgA as an auxiliary indicator for the diagnosis of nasopharyngeal carcinoma. In LMP1 (+) NPC patients, LMP1- The 30 bp deletion mutation is ubiquitous.