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在无金标准的情况下,采用贝叶斯分析评估酶联免疫吸附测定(ELISA)、间接荧光抗体试验(IFAT)和蛋白质印迹法检测猫抗婴儿利什曼原虫抗体的诊断性能。

Diagnostic performance of ELISA, IFAT and Western blot for the detection of anti-Leishmania infantum antibodies in cats using a Bayesian analysis without a gold standard.

作者信息

Persichetti Maria Flaminia, Solano-Gallego Laia, Vullo Angela, Masucci Marisa, Marty Pierre, Delaunay Pascal, Vitale Fabrizio, Pennisi Maria Grazia

机构信息

Istituto Zooprofilattico Sperimentale della Sicilia, A. Mirri, Via G. Marinuzzi 3, Palermo, 90129, Italy.

Departament de Medicina i Cirurgia Animals. Facultat de Veterinària, Universitat Autònoma de Barcelona, Bellaterra, Cerdanyola, 08193, Barcelona, Spain.

出版信息

Parasit Vectors. 2017 Mar 13;10(1):119. doi: 10.1186/s13071-017-2046-3.

Abstract

BACKGROUND

Anti-Leishmania antibodies are increasingly investigated in cats for epidemiological studies or for the diagnosis of clinical feline leishmaniosis. The immunofluorescent antibody test (IFAT), the enzyme-linked immunosorbent assay (ELISA) and western blot (WB) are the serological tests more frequently used. The aim of the present study was to assess diagnostic performance of IFAT, ELISA and WB to detect anti-L. infantum antibodies in feline serum samples obtained from endemic (n = 76) and non-endemic (n = 64) areas and from cats affected by feline leishmaniosis (n = 21) by a Bayesian approach without a gold standard.

METHODS

Cut-offs were set at 80 titre for IFAT and 40 ELISA units for ELISA. WB was considered positive in presence of at least a 18 KDa band. Statistical analysis was performed through a written routine with MATLAB software in the Bayesian framework. The latent data and observations from the joint posterior were simulated in the Bayesian approach by an iterative Markov Chain Monte Carlo technique using the Gibbs sampler for estimating sensitivity and specificity of the three tests.

RESULTS

The median seroprevalence in the sample used for evaluating the performance of tests was estimated at 0.27 [credible interval (CI) = 0.20-0.34]. The median sensitivity of the three different methods was 0.97 (CI: 0.86-1.00), 0.75 (CI: 0.61-0.87) and 0.70 (CI: 0.56-0.83) for WB, IFAT and ELISA, respectively. Median specificity reached 0.99 (CI: 0.96-1.00) with WB, 0.97 (CI: 0.93-0.99) with IFAT and 0.98 (CI: 0.94-1.00) with ELISA. IFAT was more sensitive than ELISA (75 vs 70%) for the detection of subclinical infection while ELISA was better for diagnosing clinical leishmaniosis when compared with IFAT (98 vs 97%).

CONCLUSIONS

The overall performance of all serological techniques was good and the most accurate test for anti-Leishmania antibody detection in feline serum samples was WB.

摘要

背景

在猫身上,抗利什曼原虫抗体正越来越多地用于流行病学研究或猫利什曼病临床诊断。免疫荧光抗体试验(IFAT)、酶联免疫吸附测定(ELISA)和蛋白质印迹法(WB)是最常用的血清学检测方法。本研究旨在采用无金标准的贝叶斯方法,评估IFAT、ELISA和WB检测从流行地区(n = 76)和非流行地区(n = 64)采集的猫血清样本以及感染猫利什曼病的猫(n = 21)血清中抗婴儿利什曼原虫抗体的诊断性能。

方法

IFAT的临界值设定为滴度80,ELISA的临界值设定为40个ELISA单位。当存在至少一条18 kDa条带时,WB被视为阳性。通过在贝叶斯框架下使用MATLAB软件编写的程序进行统计分析。在贝叶斯方法中,使用吉布斯采样器通过迭代马尔可夫链蒙特卡罗技术模拟联合后验的潜在数据和观测值,以估计三种检测方法的敏感性和特异性。

结果

用于评估检测性能的样本中,血清阳性率中位数估计为0.27[可信区间(CI)=0.20 - 0.34]。三种不同方法的敏感性中位数分别为:WB为0.97(CI:0.86 - 1.00),IFAT为0.75(CI:0.61 - 0.87),ELISA为0.70(CI:0.56 - 0.83)。WB的特异性中位数达到0.99(CI:0.96 - 1.00),IFAT为0.97(CI:0.93 - 0.99),ELISA为0.98(CI:0.94 - 1.00)。在检测亚临床感染时,IFAT比ELISA更敏感(75%对70%),而在诊断临床利什曼病方面,ELISA比IFAT表现更好(98%对97%)。

结论

所有血清学技术的总体性能良好,在猫血清样本中检测抗利什曼原虫抗体最准确的检测方法是WB。

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