双温聚合酶链反应检测EB病毒

Detection of Epstein-Barr virus DNA by two temperature PCR cycles

应用双温循环ＰＣＲ（聚合酶链反应）方法检测鼻咽癌石蜡包埋组织、活检或冰冻组织、患者鼻咽拭子或血液中的ＥＢ病毒（Ｅｐｓｔｅｉｎ－Ｂａｒｒｖｉｒｕｓ）ＤＮＡ。模板ＤＮＡ制备方法简便，无需酚提取步骤；双温ＰＣＲ快速（约需１．５小时）、灵敏。目前已用于临床检验。

pstein-Barr virus(EBV)DNA was examined by two temperature polymerase cbain reaction cycles in nasopharyngeal carcinoma(NPC) paraffin-embedded and biopsy tissue,nasopharyngeal swab andblood derived from patients with NPC。All the samples were treated with proteinase K lysis buffer at 55℃ for three hours and it was not necessary to purify DN A by phenol-chloroform-isoamyl alcohol ex-traction。Two temperature PCR cycles consisted of l min at 94℃ followed by l min at 68℃ and thesetwo cycles were repeated for 30 times, Then the samples were kept at 68℃ for 7 min。The whole proce-dure took about l.5 hours.The amplified product l2l bp DNA, was examined by Nusieve 3:l agarose(2%～3%).This method may be used for supplementary diagnosis of Burkitt's lymphoma,infectionsmononucleosis and Hodgkin's lymphoma etc.