鼻咽癌病人外周血EBV DNA最低检出限与不同核酸抽提试剂关系的分析

Relationship between the minimum detection limit of peripheral blood EBV-DNA and different nucleic acid extraction kits in NPC patients

目的 探讨两种核酸提取方法与鼻咽癌患者外周血EBV DNA最低检出限的关系。 方法 收集40例EB病毒感染鼻咽癌患者全血,分别采用核酸释放剂法与柱提法抽提DNA,检测EBV DNA载量,比较两种核酸提取方法检测的检出率,并确认其对应最低检出限。 结果 40例携带EBV鼻咽癌患者全血,使用核酸释放剂法检测EBV DNA其检出率为90.0%明显高于柱提法的检出率62.6%,差异有统计学意义（χ^2=8.352,P=0.008）。按国家行业标准验证的核酸释放剂法检测最低检出限为400 copies/ml;采用柱提法最低检出限为900 copies/ml。 结论 选用核酸抽提试剂检测鼻咽癌病人外周血EBV DNA时,所选择试剂应该满足或优于EBV DNA试剂盒最低检测限的性能要求。选用合适的核酸抽提试剂与EBV DNA检测试剂能够客观的反映鼻咽癌患者体内EB病毒的变化,为临床的诊断及治疗提供准确可靠的数据。

Objective To explore the relationship of two different nucleic acid extraction methods with the minimum detection limit of Epstein-Barr virus （EBV） DNA in peripheral blood of patients with nasopharyngeal carcinoma （NPC）. Methods The nucleic acid release agent method and spin columns method were used to measure EBV-DNA load in 40 peripheral blood specimens from NPC patients, their detection rates were compared, and their minimum detection limits were identified. Results Among the 40 whole blood samples detected, the EBV-DNA-positive detection rate of the nucleic acid release agent method was significantly higher than that of the spin columns method, with a statistically significant difference （90.0% vs. 62.6%, χ^2=8.352, P=0.008）. The national professional standard verified that the minimum detection limits of EBV-DNA load by the nucleic acid release agent method and spin columns method were 400 copies/ml and 900 copies/ml respectively. Conclusions Quantification of EBV in peripheral blood of NPC patients should use the nucleic acid extraction kit by which the requirement of the minimum detection limit can be reached or surpassed. The appropriate reagent canObjectively reflect the change of EBV in the blood, and provide exact and reliable data for clinical diagnosis and treatment of EBV-associated NPC.