【摘要】 目的研究人乳头瘤病毒(HPV)11型 E7蛋白在原核细胞的表达及其在尖锐湿疣(CA)血清学诊断中的应用价值。方法用 PCR 从 CA 患者组织中扩增 HPV11 E7全长基因,构建重组质粒 pET32a(+)/HPV11 E7,经异丙基硫代-β-D-半乳糖苷(IPTG)诱导后表达,用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳和蛋白质印迹法(WB)分析鉴定;经镍螯合亲和层析胶体(Ni-NTA Agarose)纯化 HPV11 E7融合蛋白后作抗原,用间接 ELISA 法对93例 CA 患者、43例宫颈癌患者和58名健康对照者血清标本进行 IgG 抗体检测。结果 HPV11 E7融合蛋白在原核表达系统中呈较高效表达(浓度为40μg/ml)。CA 组、宫颈癌组和健康对照组血清 IgG 抗体均值分别为1.545±0.131、0.586±0.155和0.674±0.150,阳性率分别为76.3%(71/93)、11.6%(5/43)和5.2%(3/58);CA 组血清 IgG抗体均值及阳性率均高于宫颈癌组和对照组(均 P<0.01)。结论 HPV11 E7融合蛋白具较强的抗原性,用于 CA 血清学诊断试剂的研制具有临床应用价值。更多还原

【Abstract】 Objective To investigate expression of Human papillomavirus (HPV) 11 type E7 protein antigen in prokaryotic cells and its potential use for the serodiagnosis of condyloma acuminatum (CA).Methods The full-length gene encoding for HPV11 E7 protein was amplified by PCR,and cloned into vector pET32a(+) to form recombinant pET32a(+)/HPVll E7 plasmid.The fusion His-E7 protein was expressed and analyzed by using SDS-PAGE and Western blotting.Using ELISA assay,HPV11 E7 fusion protein were also used to screen human serum IgG antibody from 93 patients with CA,43 patients with cervix cancer and 58 healthy control subjects.Results Highly expressed fusion His-E7 protein was obtained,and purified protein served as a special diagnostic antigen to screen human serum antibody for CA serodiagnosis.It showed that CA group,cervix cancer group and healthy control human serum IgG antibody average value were 1.545±0.131,0.586±0.155 and 0.674±0.150 respectively,positive rate were 76.3% (71/93),11.6% (5/43) and 5.2% (3/58).There was significantly difference between the CA group to compare cervix cancer group and healthy control (P<0.01).Conclusion It is likely that HPV11 E7 fusion protein could be used as a potential antigen for CA serodiagnosis.更多还原