Minas A, Stournara A, Minas M, Stack J, Petridou E, Christodoulopoulos G, Krikelis V
Technological Educational Institution of Larissa, Faculty of Health Professions, Laboratory of Microbiology, Larissa, 411 10, Greece.
J Immunol Methods. 2007 Mar 30;320(1-2):94-103. doi: 10.1016/j.jim.2006.12.008. Epub 2007 Jan 16.
Fluorescence polarization assay (FPA) is a relatively new test for the serological diagnosis of Brucella spp. infection in animals. FPA, carried out in 96-well microplate format, was validated here for diagnosing B. melitensis infection in sheep and goats. This study included sera from 1933 sheep and goats, from animals reared in naturally infected flocks (verified by culture) and showing a positive reaction to two different tests conducted in parallel. In addition, 2154 sera originating from healthy sheep and goats, reared in areas where B. melitensis had never been isolated, were assayed. The optimum cut-off value offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 15 mP over the mean value of the buffer control used in each microplate as determined by receiver operating characteristic analysis. The DSn and DSp of the FPA for small ruminants carried out in microplates at this cut-off value were calculated to be 95.9% and 97.9% with 95% confidence intervals (95% CI) of 94.9-97.7% and 97.2-98.4%, respectively. The accuracy of the FPA, as expressed by determination of the area under the curve, was 0.991. Indirect ELISA and FPA tests offered the highest DSn when compared with the Rose Bengal test, the complement fixation test, the modified Rose Bengal test and competitive ELISA. The parallel or serial combination of FPA with indirect ELISA offers the highest DSn and DSp. As temperature can affect the results of the FPA, all reagents must be at the same temperature and the standard for comparison must always be read under the same conditions as the sera under test. FPA performed in microplates is a promising assay; the DSn and accuracy are better than those of the tests currently approved for diagnosing B. melitensis in small ruminants. Because of its simplicity, speed, and accuracy, this test can improve capacity for laboratory testing and the efficacy of an eradication program based on a test-and-slaughter policy.
荧光偏振分析（FPA）是一种相对较新的用于动物布鲁氏菌属感染血清学诊断的检测方法。以96孔微孔板形式进行的FPA在此处经过验证，可用于诊断绵羊和山羊的羊布鲁氏菌感染。本研究纳入了1933份绵羊和山羊血清，这些血清来自自然感染羊群中饲养的动物（经培养验证），并且对同时进行的两种不同检测呈阳性反应。此外，还检测了来自从未分离出羊布鲁氏菌地区的2154份健康绵羊和山羊血清。通过受试者工作特征分析确定，在每个微孔板中使用的缓冲液对照平均值之上15 mP处，获得了最高诊断敏感性（DSn）和诊断特异性（DSp）的最佳临界值。在此临界值下，微孔板中对小型反刍动物进行的FPA的DSn和DSp分别计算为95.9%和97.9%，95%置信区间（95% CI）分别为94.9 - 97.7%和97.2 - 98.4%。以曲线下面积测定表示的FPA准确性为0.991。与玫瑰红试验、补体结合试验、改良玫瑰红试验和竞争ELISA相比，间接ELISA和FPA检测具有最高的DSn。FPA与间接ELISA的平行或串联组合提供了最高的DSn和DSp。由于温度会影响FPA的结果，所有试剂必须处于相同温度，并且用于比较的标准必须始终在与待测血清相同的条件下读取。在微孔板中进行的FPA是一种有前景的检测方法；其DSn和准确性优于目前批准用于诊断小型反刍动物羊布鲁氏菌的检测方法。由于其简单、快速和准确，该检测可提高实验室检测能力以及基于检测和屠宰政策的根除计划的效力。
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