结核分枝杆菌Rv3618蛋白在大肠埃希菌中的表达、纯化及其血清学诊断  

Expression of Rv3618 Protein of Mycobacterium Tuberculosis in E. coli and There of Diagnostic Serology

作　　者：何秀云[1] 李净[2] 郝娟[1] 葛林虎[1] 赵雅贞[1] 黄香玉[1] 陈红兵[2] 何珂[2] 王艳军[1] 

机构地区：[1]中国人民解放军第309医院中心实验室,北京100091 [2]中国人民解放军第309医院结核病四科,北京100091

出　　处：《现代检验医学杂志》2010年第6期27-29,共3页Journal of Modern Laboratory Medicine

摘　　要：目的 大肠埃希菌表达系统获重组结核分枝杆菌Rv3618蛋白,并评价其用于肺结核病血清学诊断的价值.方法 大肠埃希菌表达重组Rv3618蛋白,离子和疏水层析纯化获得高纯度的重组Rv3618蛋白,ELISA检测71例肺结核患者血清重组Rv3618蛋白和重组38000蛋白抗体IgG.结果 Rv3618蛋白在大肠埃希菌中表达,从大肠埃希菌纯化获得的重组Rv3618蛋白和重组38000蛋白的ELISA诊断临界值分别为A490 nm=0.51和A 490 nm=0.45.71例肺结核患者血清,重组Rv3618蛋白抗体阳性率39.4%与重组38000蛋白抗体阳性率（52.1%）比较差异无统计学意义（P＞0.05）,而与两种重组蛋白联合检测的抗体阳性率（59.2%）比较差异有统计学意义（P＜0.05）.结论 重组Rv3618蛋白可与重组38000蛋白联合用于肺结核病患者血清诊断以提高敏感性,因此,重组Rv3618蛋白可作为结核病血清学诊断的抗原之一.Objective To evaluate the potential value of a recombinant Rv3618 （rRV3618） protein of Mycobacterium tuberculosis as diagnostic serology. Methods The gene encoding a Rv3618 protein was amplified by PCR,cloned into PET-28a, and expressed in E. coll. The rRv3618 protein was purified by ion and hydrophobic chromatography. ELISA was used to detect specific IgG of rRv3618 and recombinant 38000 （r38000） proteins in sera from patients with pulmonary tuberculosis and health controls. Results The gene encoding a Rv3618 protein was expressed in E. coli. The purity of Rv3618 obtained by ion and hydrophobic chromatography was 90%. The cutoff of ELISA using tRy3618 and r38000 proteins was A490nm- 0.51 and A490nm=0.45,respectively. For 71 patients with pulmonary tuberculosis,there was no difference in the sensitivity between the rRv3618 protein （39.4%） and the r38000 protein （52.1%）,but the sensitivity of the tRy3618 protein was significantly lower than that of the tRy3618 combination with r38000 proteins （59.2%）.Conclusion The joint use of the rRv3618 and r38000 proteins increased the sensitivity of diagnostic serology,and the Rv3618 protein of Mycobacteriurn tuberculosis may be among candidates as diagnostic serology.

关 键 词：重组Rv3618蛋白 肺结核 血清学诊断 

分 类 号：R378.21[医药卫生—病原生物学] Q781[医药卫生—基础医学]