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三种不同原理的间接酶联免疫吸附试验方法在严重急性呼吸道综合征诊断中的比较研究

Comparative Studies on Three Different Indirect ELISA Methods With Different Principles in the Diagnosis of SARS
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摘要 目的比较不同原理的间接酶联免疫吸附试验(ELISA)方法在严重急性呼吸道综合征(Severeacuterespirato-rysyndrome,SARS)诊断中的应用,以便研制开发更先进的诊断试剂,应对可能发生的SARS再次流行。方法对山西省疾病预防控制中心(CDC)送检的正常成年人血清标本44份,首都医科大学附属宣武医院住院SARS患者血清标本62份,宁夏回族自治区CDC送检SARS患者及疑似患者血清28份,分别采用三种不同原理的间接ELISA试剂进行抗体检测:①美国CDC采用全病毒灭活抗原包被,检测血清IgA、IgM、IgG抗体的试剂;②国内某公司采用全病毒灭活抗原包被,检测血清IgG及IgM抗体试剂;③基因工程表达并纯化刺突蛋白(spikeprotein,S蛋白)和核衣壳蛋白(nucleocapsidprotein,N蛋白)抗原包被,检测血清中IgM及IgG抗体的试剂。结果通过比较研究显示:试剂①与试剂③的特异性较高,3种试剂的敏感性有较高的一致性。结论由于全病毒裂解抽提液作包被抗原其生产过程存在生物安全隐患,要求条件高,所以基因工程表达并纯化的蛋白来制备SARS冠状病毒抗体检测试剂有自身的优越性。由于N蛋白具有较强的免疫原性,在各毒株之间的变异又很小,因此以体外基因重组N蛋白代替全病毒裂解液蛋白,是SARS抗体检测试剂的发展方向。 Objective Comparing different indirect enzyme-linked immunosorbent assay(ELISA)methods with different principles in the diagnosis of severe acute respiratory syndrome (SARS)in order to develop more advanced diagnostic reagents,and deal with the possible reappearance of SARS. Methods 3 different indirect ELISA tests were used to detect 44 serum samples of healthy people collected from Shaanxi CDC,62 serum samples of SARS patients collected from Xuanwu hospital in Beijing and 28 serum samples of suspected SARS patients collected from Ningxia CDC.The three reagents are:(1)SARS coronavirus Vero E6 lysate antigen coated plate detects IgA,IgG, and IgM in serum produced by American CDC,(2) SARS coronavirus lysate antigen coated plate detects IgM and IgG separately produced by a company in our country,(3) Genetic recombinant SARS spike protein(S protein)and nucleocapsid protein(N protein)antigen coated plate detects IgM and IgG individually produced by China CDC. Results Our results show that:reagent(1)and reagent(3)have high specificities and all three reagents have almost the same sensitivities. Conclusions Because the procedure of producing SARS coronavirus lysate antigen is not very safe,the demand of the facilities and staff is strict. SARS antibody detection reagents using genetic recombinant proteins have their own advantages.N protein has strong immunogenicity and the diversity among different isolates is not so much,so genetic recombinant N protein antigen coated plate is [LM]the direction of producing new SARS antibodies detection reagent.
作者 朱贞 张燕 毛乃颖 刘中华 崔爱利 蒋小泓 陈彪 韩玉霞 詹军 吴立平 许文波
机构地区 中国疾病预防控制中心病毒病预防控制所 首都医科大学附属北京宣武医院 宁夏回族自治区疾病预防控制中心 山西省疾病预防控制中心
出处 《中国计划免疫》 2005年第3期198-203,共6页 Chinese Journal of Vaccines and Immunization
关键词 严重急性呼吸道综合征(SARS)冠状病毒 间接酶联免疫吸附试验 SARS抗体检测 比较研究 SARS coronavirus Indirect ELISA SARS antibodies detection Comparative studies
分类号 R373 [医药卫生—病原生物学]
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中国计划免疫
2005年 第3期

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