南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (11): 1747-1751.doi: 10.12122/j.issn.1673-4254.2021.11.22

• • 上一篇    

登革病毒IgG抗体的检测：基于登革病毒E蛋白的荧光素酶免疫吸附法

刘金月，李晓霞，王海鹰，唐时幸，万成松   

  1. 南方医科大学公共卫生学院，广东 广州 510515
  • 出版日期:2021-11-20 发布日期:2021-12-10

Dengue virus E protein-based luciferase immunosorbent assay for detecting dengue virus IgG antibody

LIU Jinyue, LI Xiaoxia, WANG Haiying, TANG Shixing, WAN Chengsong   

  1. School of Public Health, Southern Medical University, Guangzhou 510515, China
  • Online:2021-11-20 Published:2021-12-10

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摘要： 目的 建立基于登革病毒E蛋白特异性抗原的荧光素酶免疫吸附法（DENV-LISA），用于检测DENV IgG抗体。方法 分别构建DENV1-E1、DENV2-E2与荧光素酶的融合表达质粒，转染293T细胞后获得含特异性抗原与荧光素酶的融合蛋白，建立DENV-LISA，评价其特异度和灵敏度，并与商用登革病毒IgG抗体检测试剂盒比较。结果 DENV-LISA阳性检出率32.4%，特异度96.6%，商用检测试剂盒阳性检出率35.3%，差异无统计学意义（P>0.05）；阳性样本稀释6400倍可判断阳性，灵敏度高；同批板间、板内检测值差异无统计学意义（P>0.05）；重复性良好。结论 建立基于登革病毒E蛋白的荧光素酶免疫吸附法，对登革病毒IgG抗体有较高特异度和灵敏度，可用于登革病毒感染的早期筛查、监测预警、流行病学调查等。

关键词: 登革病毒；荧光素酶；E蛋白；IgG抗体；检测

Abstract: : Objective To establish a luciferase immunosorbent assay (DENV-LISA) based on dengue virus (DENV) E protein, a specific antigen of DENV, for detection of DENV IgG antibody. Methods The fused expression plasmids of DENV1-E1 and DENV2-E2 with luciferase were constructed. The plasmids were transfected into 293T cells, and the fusion protein containing the specific antigen and luciferase was obtained for establishing DENV-LISA. The specificity and sensitivity of DENV-LISA were assessed and compared with those of commercial DENV IgG antibody detection kit (ELISA). Results The established DENV-LISA had a positive detection rate of 32.4% and a specificity of 96.6%, showing a similar positive detection rate with the commercial ELISA kit (35.3%; P>0.05). DENV-LISA was capable of detecting positive samples with a 1: 6400 dilution with a high sensitivity. The test values of DENV-LISA did not differ significantly between plates or within plates in the same batch (P> 0.05), suggesting a good reproducibility of the test. Conclusion The luciferase immunosorbent assay based on DENV E protein has high specificity and sensitivity for detecting DENV IgG antibody, and can be used for early screening, surveillance and epidemiological investigation of DENV infection.

Key words: dengue virus; luciferase; E protein; IgG antibody; detection