| 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 |
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| 引用本文： | 刘金月,李晓霞,王海鹰,唐时幸,万成松.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[J].南方医科大学学报,2021,41(11):1747-1751. |
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| 作者姓名： | 刘金月 李晓霞 王海鹰 唐时幸 万成松 |
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| 作者单位： | 南方医科大学公共卫生学院,广东 广州 510515 |
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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。
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| 关 键 词： | 登革病毒 荧光素酶 E蛋白 IgG抗体 检测 |
Dengue virus E protein-based luciferase immunosorbent assay for detecting dengue virus IgG antibody |
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| LIU Jinyue,LI Xiaoxia,WANG Haiying,TANG Shixing,WAN Chengsong.Dengue virus E protein-based luciferase immunosorbent assay for detecting dengue virus IgG antibody[J].Journal of Southern Medical University,2021,41(11):1747-1751. |
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| Authors: | LIU Jinyue LI Xiaoxia WANG Haiying TANG Shixing WAN Chengsong |
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| Affiliation: | School of Public Health, Southern Medical University, Guangzhou 510515, China |
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| 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. |
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| Keywords: | dengue virus luciferase E protein IgG antibody detection |
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