Cabana A L, Xavier M O, Mendes J F, Teles A J, Martins A M, Silva-Filho R P, Meireles M C A
Programa de Pós-graduação em Veterinária, Centro de Pesquisa e Diagnóstico em Micologia Veterinária - MICVET, Universidade Federal de Pelotas - UFPel, Av. Eliseu Maciel, s/n, Campus Universitário, CEP 96010-900, Capão do Leão, RS, Brasil.
Faculdade de Medicina - FAMED, Departamento de Parasitologia e Micologia, Área de Ciências da Saúde, Universidade Federal do Rio Grande - FURG, Rua Visconde de Paranaguá, 102, CEP 96200-190, Rio Grande, RS, Brasil.
Braz J Biol. 2019 Apr-Jun;79(2):169-173. doi: 10.1590/1519-6984.171140. Epub 2018 Aug 20.
Even today, an effective diagnostic test for aspergillosis in penguins is unknown, being the gold standard post-mortem examinations. The fungal antigen galactomannan (GM) has been used as a biomarker of disease in humans and is detected by the Platelia Aspergillus EIA (BioRad)®, a commercial kit based on the sandwich ELISA technique. It is standardized for use in neutropenic patients, however studies have demonstrated its usefulness also possible for birds. The aim of our study was to evaluate the effectiveness of Platelia Aspergillus EIA® test (BioRad-US) in the diagnosis of aspergillosis in Magellanic penguins, determining sensitivity, specificity, and positive and negative predictive values for different cut-off points. Were included in the study, blood serum samples (n = 29) Magellanic penguins in captivity that died by aspergillosis. Detection of GM was performed following manufacturer's instructions and the GM index was obtained by dividing the average value of OD of the duplicate of the clinical sample by duplicate OD of the average value of the cut-off sample provided by the kit. Through information database results were obtained for the presence of anti-Aspergillus fumigatus antibodies detected by agar gel immunodiffusion (AGID) for all serum samples. Results were analyzed using chi-square test and Kruskal-Wallis from SPSS 20.0, IBM®. ROC curve was obtained and from this, rates of sensitivity, specificity, positive and negative predictive values were also calculated based on four different cutoff points (0.5, 1.0, 1.5 and 2.0). The serum GM index did not differ between animals of the case and control group (pkw =0.097). In determining the ROC curve for serum GM detection the value of area under the curve was 0.635. From the values determined by the coordinate of the curve, four different cut points (0.5, 1.0, 1.5 and 2.0) were analyzed, resulting in sensitivity rates ranging from 86.2 to 34.5% % and specificity between 87% and 26.1%. By comparing the serum GM index in group case as the presence or absence of antibodies detected by AGID was found p=0.503. The detection of GM the Platelia Aspergillus EIA® test seems is not be useful for the diagnosis of aspergillosis in naturally infected penguins.
即使在今天，用于企鹅曲霉菌病的有效诊断测试仍然未知，死后检查是金标准。真菌抗原半乳甘露聚糖（GM）已被用作人类疾病的生物标志物，并通过曲霉菌平板酶免疫测定法（Platelia Aspergillus EIA）（伯乐公司）检测，这是一种基于夹心酶联免疫吸附测定技术的商业试剂盒。它已标准化用于中性粒细胞减少症患者，然而研究表明其对鸟类也可能有用。我们研究的目的是评估曲霉菌平板酶免疫测定法（Platelia Aspergillus EIA）（美国伯乐公司）在麦哲伦企鹅曲霉菌病诊断中的有效性，确定不同临界值的敏感性、特异性以及阳性和阴性预测值。纳入研究的有圈养的因曲霉菌病死亡的麦哲伦企鹅的血清样本（n = 29）。按照制造商的说明进行GM检测，GM指数通过将临床样本重复孔的OD平均值除以试剂盒提供的临界样本平均值的重复孔OD值来获得。通过信息数据库获得所有血清样本通过琼脂凝胶免疫扩散法（AGID）检测到的抗烟曲霉抗体的存在情况的结果。使用IBM® SPSS 20.0中的卡方检验和克鲁斯卡尔 - 沃利斯检验分析结果。获得ROC曲线，并据此基于四个不同的临界值（0.5、1.0、1.5和2.0）计算敏感性、特异性、阳性和阴性预测值。病例组和对照组动物的血清GM指数没有差异（pkw = 0.097）。在确定血清GM检测的ROC曲线时，曲线下面积值为0.635。根据曲线坐标确定的值，分析了四个不同的切点（0.5、1.0、1.5和2.0），敏感性率在86.2%至34.5%之间，特异性在87%至26.1%之间。通过比较病例组中血清GM指数与AGID检测到的抗体的有无，发现p = 0.503。曲霉菌平板酶免疫测定法（Platelia Aspergillus EIA）检测GM似乎对自然感染企鹅的曲霉菌病诊断没有用处。
