应用FQ-PCR技术测定鼻咽癌患者血浆EB病毒DNA定量分析及其临床意义

EB Virus DNA Quantitative Analysis of Plasma and Its Clinical Significance in Patients with Nasopharyngeal Application FQ-PCR Assay Technique

目的应用FQ-PCR技术测定鼻咽癌患者血浆中游离EB病毒DNA拷贝,探讨血浆EB病毒DNA定量测定的临床意义。方法选取132例鼻咽癌患者,取其外周血样本,其中84例治疗前血样,48例治疗后血样(放疗或加化疗)。另收集60例健康血样作为正常对照。使用广州中山医科大学达安基因诊断中心提供(Cat.#DA-061)的EB病毒DNA-PCR试剂盒,测定鼻咽癌患者血浆中游离EB病毒DNA拷贝,阴性对照为空白PCR反应液,阳性对照为10~6、10~4、10~2拷贝/ul的阳性模板。结果鼻咽癌组治疗前84例样本,阳性率为67.86%,鼻咽癌组治疗后48例样本阳性率为35.42%,正常对照者30例样本阳性率仅为8.33%,鼻咽癌组治疗前血浆游离EB病毒DNA的阳性率显著高于对照组,差异显著(χ~2=11.497,P=0.001);鼻咽癌组治疗后与对照组血浆游离EB病毒DNA的阳性率比较,差异较显著(χ~2=6.782,P=0.018);鼻咽癌组治疗前血浆中游离EB病毒-DNA阳性率约是治疗后的2倍,差异显著(χ~2=6.271,P=0.023)。鼻咽癌组治疗前血浆游离EB病毒DNA拷贝中位数为522.0 copies/ml,治疗后中位数为0.0,对照组中位数为0.0,鼻咽癌组治疗前的血浆游离EB病毒DNA拷贝数显著高于治疗后,两者差异具有统计学意义(U=350.0,P=0.029),而且与正常对照组拷贝数比较,差异亦显著(U=274.0,P=0.001)。Ⅰ~Ⅱ期患者的血浆游离EB病毒DNA水平显著低于Ⅲ~Ⅳ期患者(U=141.0,P=0.039)。N_0+N_1期患者的血浆EB病毒DNA水平显著低于N_2+N_3期患者(U=147.0,P=0.031)。结论 FQ-PCR技术具有快速、精确和高灵敏性的特点,比其它传统检测手段更实用。血浆EB病毒DNA的定量PCR分析对鼻咽癌的筛选检查具有应用价值。

Objective To study the FQ-PCR technique in patients with nasopharyngeal carcinoma plasma free EB virus DNA copies,and clinical significance of plasma EB virus DNA quantitative determination .Methods 132 patients with nasopha-ryngeal carcinoma were selected , whichever peripheral blood samples , including 84 cases of blood before treatment , after treat-ment,48 cases of blood（radiotherapy or chemotherapy ）.Another 60 healthy blood samples were collected as normal controls .Tat gene diagnosis center （ Cat.#DA-061） of EB virus DNA-PCR kit plasma of patients with nasopharyngeal carcinoma of free copies of EB virus DNA negative control was blank PCR reaction solution ,a positive control of 106,104,102 copies/ul positive template. Application of statistical analysis software SPSS 20.0 experimental data processing .Results The treatment group 84 cases of na-sopharyngeal carcinoma samples ,the positive rate was 67.86%,after the treatment of 48 cases of nasopharyngeal carcinoma sam-ple positive rate was 35.42%,30 cases of normal controls samples positive rate was 8.33%,nasopharyngeal carcinoma before treatment plasma free EB virus DNA positive rate was significantly higher ,the difference was significant （χ2 =11.497,P =0.001）;compared with the control group ,plasma free EB virus DNA positive rate of nasopharyngeal carcinoma after treatment ,the 2 group was more significant （χ2 =6.782,P=0.018）;nasopharyngeal group therapy before bE viral-DNA positive rate of plasma free about 2 times after treatment,the difference was significant（χ2 =6.271,P=0.023）.Before treatment nasopharyngeal EB vi-rus DNA in plasma free copy of median 522.0copies/ml,median was 0.0 after treatment,the median of the control group was 0.0,the copy number of free EB virus DNA in plasma of nasopharyngeal carcinoma group before treatment significantly higher than after treatment,the difference was statistically significant （μ=350.0,P=0.029）,and compared with the control group ,the number of copies,the difference was also significant （μ=274.0,P=0.001）.Plasma free EB virus DNA level Ⅰ~Ⅱpatients＆amp;nbsp;was significantly lower than the Ⅲ~Ⅳpatients（μ=141.0,P=0.039）.N0 ＋N1 plasma EB virus DNA levels were significantly lower than patients with N2 ＋N3（μ=147.0,P=0.031）.Conclusion FQ-PCR technique is fast,accurate and highly sensitive, it is more practical than other traditional means of detection .Quantitative PCR analysis of EB virus DNA in plasma of nasopharyn-geal cancer screening has value .