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目的探讨网状分枝扩增法(RAM)检测非霍奇金淋巴瘤(NHL)外周血EB病毒基因在EB病毒感染与NHL发病的相关性研究中的意义。方法用人工合成EBER-1启动子基因,建立网状分枝扩增检测法,以EB病毒阳性细胞株Raji和EB病毒阴性的人白血病细胞株NB4作对照,检测120例NHL患者外周血中EB病毒基因。结果RAM最少能够检出样品中10拷贝/μl的EB病毒DNA,与多重聚合酶链反应(mPCR)的灵敏度一致;并从120例NHL患者中,检出76例阳性(63·3%),与mPCR方法的阳性检出符合率为100%。结论RAM法具有高度敏感度和特异性、操作简便,且可在等温条件下扩增核酸;同时证明NHL发病与EB病毒感染具有较高的相关性。
Objective To investigate the significance of detecting the Epstein - Barr virus (EBV) Epstein - Barr virus (Epstein - Barr virus) gene in peripheral blood of non - Hodgkin ’s lymphoma (NHL) by reticulated - branch amplification assay (RAM) Methods EBER-1 promoter gene was synthesized and the reticulocyte proliferation assay was used to detect Epstein-Barr virus positive cell line Raji and Epstein-Barr virus-negative human leukemia cell line NB4 as control. The levels of EB in 120 patients with NHL Virus gene. Results The RAM was able to detect at least 10 copies / μl of Epstein-Barr virus DNA in the sample with the same sensitivity as the multiplex polymerase chain reaction (mPCR). Of the 120 NHL patients, 76 were positive (63.3%), The positive detection rate with mPCR method was 100%. Conclusion The RAM method is highly sensitive and specific, easy to operate and can amplify nucleic acid under isothermal condition. Meanwhile, it is proved that the incidence of NHL is highly correlated with Epstein-Barr virus infection.