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用于检测犬血清中犬埃立克体抗体的间接免疫荧光试验、蛋白质印迹分析及市售酶联免疫吸附测定的比较

Comparison of an indirect immunofluorescence assay, western blot analysis, and a commercially available ELISA for detection of Ehrlichia canis antibodies in canine sera.

作者信息

O'Connor Thomas P, Hanscom Jancy L, Hegarty Barbara C, Groat Randall G, Breitschwerdt Edward B

机构信息

Department of Research and Development, IDEXX Laboratories Inc, 1 IDEXX Dr, Westbrook, ME 04092, USA.

出版信息

Am J Vet Res. 2006 Feb;67(2):206-10. doi: 10.2460/ajvr.67.2.206.

Abstract

OBJECTIVE

To examine the correlation between results for an indirect immunofluorescence assay (IFA) that uses Ehrlichia canis antigen as a substrate (ie, E canis-IFA), 2 western blot (WB) analyses, and a commercially available ELISA in the detection of E canis antibody in dog sera.

SAMPLE POPULATION

54 canine serum samples that were reactive on E canis-IFA and 16 canine serum samples that were E canis-IFA nonreactive.

PROCEDURE

Serum samples were evaluated by use of 2 WB analyses and a commercially available ELISA. Correlation between results of the 3 testing modalities (ie, IFA, WB analyses, and the ELISA) was examined by use of nonreactive (E canis-IFA reciprocal titer, < 20), low-titer (reciprocal titer, 80 to 160), medium-titer (reciprocal titer, 320 to 2,560), and high-titer (reciprocal titer, 5,120 to > 20,480) serum samples.

RESULTS

For all serum samples in the nonreactive (n = 16), medium-titer (17), and high-titer (18) groups, correlation of results among IFA, WB analyses, and the commercially available ELISA was excellent. A poor correlation was found between IFA results and those of WB analyses and the ELISA for serum samples in the low-titer group (19), with only 4 of the 19 serum samples having positive results on both WB analyses and the commercially available ELISA.

CONCLUSIONS AND CLINICAL RELEVANCE

The discrepancy between E canis-IFA, WB analyses, and the commercially available ELISA results for the low-titer serum samples may be related to a high IFA sensitivity or, more likely, a lack of specificity associated with cross-reactivity among Ehrlichia spp.

摘要

目的

检测以犬埃立克体抗原为底物的间接免疫荧光试验(IFA,即犬埃立克体 - IFA)结果、两种蛋白质印迹法(WB)分析结果以及一种市售酶联免疫吸附测定(ELISA)在检测犬血清中犬埃立克体抗体时的相关性。

样本群体

54份在犬埃立克体 - IFA检测中呈阳性反应的犬血清样本以及16份在犬埃立克体 - IFA检测中呈阴性反应的犬血清样本。

方法

采用两种蛋白质印迹法分析和一种市售酶联免疫吸附测定对血清样本进行评估。通过使用阴性反应(犬埃立克体 - IFA效价倒数,<20)、低滴度(效价倒数,80至160)、中滴度(效价倒数,320至2560)和高滴度(效价倒数,5120至>20480)血清样本,检测三种检测方法(即IFA、蛋白质印迹法分析和ELISA)结果之间的相关性。

结果

在阴性反应组(n = 16)、中滴度组(17份)和高滴度组(18份)的所有血清样本中,IFA、蛋白质印迹法分析和市售ELISA结果之间的相关性极佳。在低滴度组(19份)血清样本中,IFA结果与蛋白质印迹法分析和ELISA结果之间的相关性较差,19份血清样本中只有4份在蛋白质印迹法分析和市售ELISA中均呈阳性结果。

结论及临床意义

低滴度血清样本的犬埃立克体 - IFA、蛋白质印迹法分析和市售ELISA结果之间的差异可能与IFA的高敏感性有关,或者更有可能与埃立克体属之间的交叉反应性导致的缺乏特异性有关。

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