酶斑点杂交法与双抗体夹心法检测汉坦病毒感染的比较  

Comparison of the enzyme dot hybridization assay with double antibody sandwich assay in the detection of Hantavirus infection

作　　者：刘兵[1] 史俊岩[1] 王舰[1] 王美莲[1] 王斯[1] 郑兰艳[1] 牟玲[2] 罗恩杰[1] 

机构地区：[1]中国医科大学病原生物教研室,沈阳110001 [2]沈阳市传染病院出血热研究所

出　　处：《中国人兽共患病学报》2009年第6期531-533,共3页Chinese Journal of Zoonoses

基　　金：辽宁省自然科学基金资助项目(20062101)

摘　　要：目的将酶标抗汉坦病毒NP抗原单链抗体应用于酶斑点杂交法和双抗体夹心法,检测汉坦病毒感染,探索价格低廉早期HFRS诊断试剂。方法以戊二醛偶联辣根过氧化物酶和抗汉坦病毒NP抗原单链抗体,制备酶标单链抗体,并应用于酶斑点杂交法和双抗体夹心法中,检测56份早期HFRS患者血清及66份阴性对照血清。结果56份HFRS患者血清中,双抗体夹心法检测出29例阳性,阳性率为51.79%,酶斑点杂交法检测出28例阳性,阳性率为50.00%,两方法检测66例阴性对照血清,结果均为阴性。统计学处理,两方法差异不显著。结论以酶标抗汉坦病毒NP抗原单链抗体制备的酶斑点杂交和双抗体夹心诊断试剂,均为稳定性好、假阳性率低、造价低廉、诊断可靠的诊断试剂。Peroxidase horseradish （HRP）-labeled single chain fragment of variable region （scFv） against Hantavirus nucleocapsid protein （NP） antigen was applied in enzyme dot hybridization assay and double antibody sandwich assay to detect Hantavirus infection in order to develop the cheap diagnostic reagents for the early diagnosis of haemorrhagic fever with renal syndrome （HFRS）. In these assays, the scFv against Hantavirus NP antigen was conjugated with HRP b glutaraldehyde to prepare the HRP-labeled scFv and the HRP-labeled scFv was then used in enzyme dot hybridization and double antibody sand wich assay to detect Hantavirus infection samples of patients with early HFRS and 66 sera of healthy individuals as control. It was found that using enzyme dot hybridization assay, the positive detection rate was 50. 00% （28/29）, while that using double antibody sandwich assay is 51.78（29/56）, the serum samples of the 66 control individuals showed negative results by using both assays. The results were dealt with statistical methods and the difference in both methods was not obvious. It is concluded that the method of assays using-labeled scFv against Hantavirus NP antigen and applied in enzyme dot hybridization method and double antibody sandwich method are proved to be stable, cheap and reliable with very low false positive rate.

关 键 词：汉坦病毒 SCFV 酶斑点杂交法 双抗体夹心法 

分 类 号：R373.3[医药卫生—病原生物学]