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禽白血病病毒衣壳蛋白抗原p27快速检测免疫层析试纸条的研制与评价

Development and evaluation of an immunochromatographic strip for rapid detection of capsid protein antigen p27 of avian leukosis virus.

作者信息

Qian Kun, Liang You-zhi, Yin Li-ping, Shao Hong-xia, Ye Jian-qiang, Qin Ai-jian

机构信息

Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, Yangzhou 225009, China; Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.

Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, Yangzhou 225009, China.

出版信息

J Virol Methods. 2015 Sep 1;221:115-8. doi: 10.1016/j.jviromet.2015.04.033. Epub 2015 May 11.

Abstract

A rapid immunochromatographic strip for detecting capsid protein antigen p27 of avian leukosis virus was successfully developed based on two high-affinity monoclonal antibodies. The test strip could detect not only 600pg purified recombinant p27 protein but also quantified avian leukosis virus as low as 70 TCID50, which has comparative sensitivity to the commercial enzyme-linked immunosorbent assay (ELISA) kit. For the evaluation of this test strip, 1100 samples consisting of cloacal swabs, meconium collected from the earliest stool of one day old chicken and virus isolates were assessed both by the strip and by the commercial ELISA kit. The agreement between these two tests was 93.91%, 93.42% and 100%, respectively. The sensitivity and specificity of the strip were also calculated by using the ELISA kit as the standard. This immunochromatographic strip provides advantages of rapid and simple detection of capsid protein antigen p27 of avian leukosis virus, which could be applied as an on-site testing assay and used for control and eradication programs of avian leukosis disease.

摘要

基于两种高亲和力单克隆抗体,成功开发出一种用于检测禽白血病病毒衣壳蛋白抗原p27的快速免疫层析试纸条。该试纸条不仅能检测600pg纯化的重组p27蛋白,还能定量低至70 TCID50的禽白血病病毒,其灵敏度与市售酶联免疫吸附测定(ELISA)试剂盒相当。为评估该试纸条,采用该试纸条和市售ELISA试剂盒对1100份样本进行了检测,这些样本包括泄殖腔拭子、从1日龄雏鸡最早粪便中收集的胎粪以及病毒分离株。这两种检测方法的一致性分别为93.91%、93.42%和100%。还以ELISA试剂盒为标准计算了试纸条的灵敏度和特异性。这种免疫层析试纸条具有快速、简便检测禽白血病病毒衣壳蛋白抗原p27的优点,可作为现场检测方法,用于禽白血病疾病的防控和根除计划。

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