西方马脑炎病毒抗体快速检测试纸条的研制

中国媒介生物学及控制杂志 ›› 2011, Vol. 22 ›› Issue (5): 421-423.

西方马脑炎病毒抗体快速检测试纸条的研制

王林林^1,2, 杨宇¹, 王旺¹, 王振东², 王静¹

1. 中国检验检疫科学研究院,北京 100123;
2. 沈阳农业大学生物科学技术学院,沈阳 110866

收稿日期:2011-04-29 出版日期:2011-10-20 发布日期:2011-10-20
通讯作者: 王静,Email: wangjing0115@126.com
作者简介:王林林(1983-),女,硕士,从事媒介生物快速检测技术研究。Email: linwang2009@126.com
基金资助:
质检公益项目(2007GYJ024,2007GYJ023);国家质检总局科技计划项目(2010IK211)

Design of a test strip for rapid detection of antibodies against Western equine encephalitis virus

WANG Lin-lin^1,2, YANG Yu¹, WANG Wang¹, WANG Zhen-dong², WANG Jing¹

1. China Academy of Inspection and Quarantine, Beijing 100123, China;
2. Shenyang Agricultural University, Institute of Biological Science and Technology, Shenyang 110866, Liaoning Province, China

Received:2011-04-29 Online:2011-10-20 Published:2011-10-20
Supported by:
Supported by the Project of AQSIQ Public Benefic(No. 2007GYJ024, 2007GYJ023)and Science and Technology Projects ofAQSIQ(No. 2010IK211)

摘要/Abstract

摘要：

目的研制一种快速检测西方马脑炎病毒(WEEV)抗体的试纸条。方法利用胶体金标记和免疫层析技术,用25 nm 胶体金标记葡萄球菌A蛋白,并用硝酸纤维膜包被WEEV抗原及羊抗兔IgG作为检测带和质控带,研制出WEEV抗体快速检测试纸条。对该试纸条的敏感性、特异性和稳定性做出评价,并拟合检测曲线进行半定量检测。在健康人血清、兔血清和鼠血清中添加WEEV抗体模拟污染样品,评价该方法的检测能力。结果该试纸条可在20 min内完成定性和半定量检测,灵敏度为120 ng/ml,模拟样品的检测灵敏度也相同。线性范围120～1200 ng/ml。对发病症状相似以及近缘的其他病毒进行检测,均无非特异反应。试纸条在37 ℃下保存1个月,检测结果不变。结论该试纸条具有快速、简便、灵敏、特异、稳定的特点,适用于现场检测。

关键词: 西方马脑炎病毒, 免疫层析技术, 胶体金试纸, 定性检测, 定量检测

Abstract:

Objective To develop a test strip for rapid detection of antibodies against Western equine encephalitis virus (WEEV). Methods Staphylococcal protein A was labeled with 25 nm colloidal gold via immune-chromatography; cellulose nitrate-coated WEEV antigens and goat anti-rabbit antibodies were used as the test zone and the control zone, respectively. The sensitivity, specificity and stability of the test strip were evaluated and semi-quantitative detection was conducted with a curve of best fit. The detection capability of the test strip was assessed using healthy human serum, rabbit serum and rat serum samples containing simulated WEEV antibodies. Results The qualitative and semi-quantitative detection was completed within 20 min, with a resulting sensitivity of 120 ng/ml in simulated samples. The linear range of sensitivity was 120-1200 ng/ml. No nonspecific reaction was observed when applied to other homologous viruses and those leading to similar symptoms. Storage at 37 ℃ over a month did not make a difference in the test result. Conclusion The test strip is a fast, simple, sensitive, specific and reliable tool for detection of WEEV and is suitable for field application.

Key words: Western equine encephalitis virus, Immunochromatography, Immunogold labeling, Qualitative detection, Quantitative detection

中图分类号:

R373

王林林, 杨宇, 王旺, 王振东, 王静. 西方马脑炎病毒抗体快速检测试纸条的研制[J]. 中国媒介生物学及控制杂志, 2011, 22(5): 421-423.

WANG Lin-lin, YANG Yu, WANG Wang, WANG Zhen-dong, WANG Jing. Design of a test strip for rapid detection of antibodies against Western equine encephalitis virus[J]. Chines Journal of Vector Biology and Control, 2011, 22(5): 421-423.

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