【摘要】 目的探讨实时荧光定量PCR和RT-PCR检测HPV-16E6和E7基因的实验方法。方法将制备的含HPV-16E6和E7基因的质粒作为标准品,建立实时荧光定量PCR和RT-PCR方法,分别对3种宫颈癌细胞株进行HPV-16E6和E7基因的DNA和RNA拷贝数的定量检测。结果建立的实时荧光定量PCR标准曲线相关系数大于0.99,PCR效率在90%以上。HPV-16E6和E7基因的DNA和RNA拷贝数在宫颈癌细胞株CaSki、SiHa和HeLa中从高到低依次是CaSki>SiHa>HeLa。结论用实时荧光定量PCR和RT-PCR方法研究HPV-16E6和E7基因的DNA和RNA含量结果可靠,为进一步研究两个基因与宫颈病变的关系奠定了基础。更多还原

【Abstract】 Objective To establish a method of using real-time fluorescence quantitative PCR and RT-PCR to detect the E6 and E7 genes of human papillomavirus type 16 (HPV-16). Methods Plasmids containing HPV-16 E6 or E7 were used to generate absolute standard curves. Three cervical carcinoma cell lines CaSki, SiHa and HeLa were tested by real-time fluorescence quantitative PCR and RT-PCR analyses for the expressions of HPV-16 E6 and E7. Results The correlation coefficients of standard curves were larger than 0.99, and the PCR efficiency was more than 90%. The relative levels of HPV-16 E6 and E7 DNA and RNA were CaSki>SiHa>HeLa cell. Conclusion HPV-16 E6 and E7 quantum by real-time fluorescence quantitative PCR and RT-PCR analyses may serve as a reliable and sensitive tool. This study provides the possibility of further researches on the relationship between HPV-16 E6 or E7 copy number and cervical carcinoma.更多还原