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通过酶联免疫吸附测定法比较全病毒、N蛋白和N199蛋白对严重急性呼吸综合征冠状病毒进行快速诊断的有效性

Comparison of effectiveness of whole viral, N and N199 proteins by ELISA for the rapid diagnosis of severe acute respiratory syndrome coronavirus.

作者信息

Guo Zhong-min, Lu Jia-hai, Han Wen-yu, Liu Ze-yu, Li Guo-wei, Liao Jia-wei, Wang Shu-min, Wu Ying-song, Zheng Huan-ying, Zhong Nan-shan, Zhu Xing-quan

机构信息

Experiment Animal Center, Sun Yat-sen University, Guangzhou 510080, China.

出版信息

Chin Med J (Engl). 2007 Dec 20;120(24):2195-9.

Abstract

BACKGROUND

Although severe acute respiratory syndrome (SARS) has been controlled, the subsequently emerging sporadic cases in 2004 emphasize the necessity of developing a rapid diagnostic method, which would be of great help in clinical diagnosis and also wild host screening. This study aims to establish an effective and rapid serological tool for the diagnosis of SARS-CoV by comparison among whole viral, N and N199 proteins by ELISA.

METHODS

SARS-CoV N and N199 (a truncated nucleocapsid gene) genes were cloned, expressed, identified by Western blotting, and applied in screening of human and swine samples. Sera of SARS convalescent-phase patients, normal human sera, sera of patients with other respiratory diseases, and swine sera were screened by ELISA, with whole SARS-CoV F69, N and N199 proteins as antigens.

RESULTS

The sensitivity and specificity of N and N199 proteins in human sera diagnosis were approximate (P = 0.743), which was higher than whole viral protein but the difference was not significant (P = 0.234). The N199 protein proved to be more specific in swine sera screening than whole viral and N protein (P < 0.001).

CONCLUSION

N199 protein is feasible in both clinical diagnosis and SARS-CoV reservoir screening.

摘要

背景

尽管严重急性呼吸综合征(SARS)已得到控制,但2004年出现的散发病例强调了开发一种快速诊断方法的必要性,这将对临床诊断和野生宿主筛查有很大帮助。本研究旨在通过酶联免疫吸附测定(ELISA)比较全病毒、N蛋白和N199蛋白,建立一种有效且快速的用于诊断SARS冠状病毒的血清学工具。

方法

克隆、表达SARS冠状病毒N基因和N199基因(截短的核衣壳基因),通过蛋白质免疫印迹法进行鉴定,并应用于人和猪样本的筛查。以全SARS冠状病毒F69、N蛋白和N199蛋白为抗原,通过ELISA对SARS康复期患者血清、正常人血清、其他呼吸道疾病患者血清和猪血清进行筛查。

结果

N蛋白和N199蛋白在人血清诊断中的敏感性和特异性相近(P = 0.743),高于全病毒蛋白,但差异不显著(P = 0.234)。在猪血清筛查中,N199蛋白比全病毒蛋白和N蛋白更具特异性(P < 0.001)。

结论

N199蛋白在临床诊断和SARS冠状病毒宿主筛查中均可行。

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