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用于定量检测抗阿留申水貂病病毒抗体的酶联免疫吸附测定的准确性

Accuracy of enzyme-linked immunosorbent assays for quantification of antibodies against Aleutian mink disease virus.

作者信息

Farid A H, Rupasinghe P P

机构信息

Department of Plant and Animal Sciences, Dalhousie University Faculty of Agriculture, Truro, Nova Scotia, B2N 5E3, Canada.

Department of Plant and Animal Sciences, Dalhousie University Faculty of Agriculture, Truro, Nova Scotia, B2N 5E3, Canada.

出版信息

J Virol Methods. 2016 Sep;235:144-151. doi: 10.1016/j.jviromet.2016.06.004. Epub 2016 Jun 6.

Abstract

There is a growing interest among mink ranchers to select their stock for tolerance to the Aleutian mink disease virus (AMDV). Enzyme-linked immunosorbent assays (ELISA) are used to identify mink which have low anti-AMDV antibody titres and are expected to tolerate the AMDV infection. The objective of this study was to calculate the accuracy of three ELISA systems which were performed on blood or serum of AMDV-inoculated American mink (Neovison vison) at five laboratories in Canada, USA, Finland, the Netherlands and Denmark. The accuracy was determined by comparing the ELISA results with antibody titres measured by the counter-immunoelectrophoresis (CIEP) using 10 two-fold serial dilutions of the plasma. Antibody titres of 880 black mink which were inoculated with a spleen homogenate from a naturally infected mink were measured between 16 and 176 weeks post-inoculation. Each ELISA result from every laboratory covered a wide range of antibody titres and the Spearman's rank correlation coefficients between CIEP and ELISA results from different laboratories varied between 0.41 and 0.83, indicating a low to moderate accuracy of ELISA systems for ranking mink by antibody titre. The recombinant VP2-based ELISA used in the Netherlands and Finland ranked the mink by antibody titres more accurately than did the AMDV-G-based ELISA platforms developed in Denmark and the USA, suggesting that the source of antigen was one of the factors affecting the accuracy of ELISA results. It was concluded that the ELISA systems, particularly those based on AMDV-G antigen, require further refinement to improve their accuracy for ranking mink by antibody titre.

摘要

水貂养殖者对选择耐阿留申水貂病病毒(AMDV)的种畜越来越感兴趣。酶联免疫吸附测定(ELISA)用于识别抗AMDV抗体滴度低且有望耐受AMDV感染的水貂。本研究的目的是计算在加拿大、美国、芬兰、荷兰和丹麦的五个实验室,对接种AMDV的美国水貂(Neovison vison)的血液或血清进行的三种ELISA系统的准确性。通过将ELISA结果与使用血浆的10倍系列稀释液通过对流免疫电泳(CIEP)测量的抗体滴度进行比较来确定准确性。在接种后16至176周之间,测量了880只接种自然感染水貂脾脏匀浆的黑色水貂的抗体滴度。每个实验室的每个ELISA结果都涵盖了广泛的抗体滴度范围,不同实验室的CIEP和ELISA结果之间的斯皮尔曼等级相关系数在0.41至0.83之间,表明ELISA系统根据抗体滴度对水貂进行排名的准确性较低至中等。荷兰和芬兰使用的基于重组VP2的ELISA比丹麦和美国开发的基于AMDV-G的ELISA平台更准确地根据抗体滴度对水貂进行排名,这表明抗原来源是影响ELISA结果准确性的因素之一。得出的结论是,ELISA系统,特别是基于AMDV-G抗原的系统,需要进一步改进以提高其根据抗体滴度对水貂进行排名的准确性。

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