汉坦病毒核蛋白基因表达及产物抗原活性的研究

为了制备肾综合征出血热（ ＨＦＲＳ） 诊断用基因工程抗原，构建了合汉坦病毒７６１１８ 株编码核蛋白（ ＮＰ） 基因片段的重组质粒ｐＢＶ２２０Ｈ１－３Ｓ， 经转化大肠杆菌和诱导表达，ＳＤＳＰＡＧＥ 显示菌体中有分子量约为４８ ＫＤ 的新蛋白质生成。用经包含体纯化获得的表达产物做抗原，以间接酶联免疫（ＥＬＩＳＡ） 检测了４８ 份疑似出血热患者的血清特异性ＩｇＧ，其结果与经典的间接免疫荧光（ＩＦＡ） 检测的阴性、阳性及总符合率分别为１００ ％ 、９２－８６ ％ 和９３－７５ ％ 。证实该重组ＮＰ 具有良好的生物活性和诊断抗原价值

Expression of nucleocapsid protein gene of Hantavirus and product's antigen function

For obtaining the reombinant diagnostic antigen of Hantavirus,pBV220-H1 3S was constructed which contained gene segment coding the Nucleocapsidprotein(NP) of hantan virus strain 76118. After this plasmid was transformed to and expressed in E. coli,It was found by SDS-PAGE that a new protein emerging in the baoteria cells with the Molecular Weight of about 48KD which is consistent with that of NP. Through purification of inclusion body, the recombinant NP (rNP) was used as coating antigen in Enzyme linked Immuno- sorbent Assay, detecting the specific IgG in 48 sera of suspect patients of Haemorrhagic Fever with Renal Syndrome. The result coincided with that by indirect immuno fluorescent Assay by percentage of 100,92.88 and 93.75 for negative, positive and all samples respectively.The data indicated that this rNP was effective and valuable for use as diagnostic antigen.