Roussel Sandrine, Reboux Gabriel, Rognon Bénédicte, Monod Michel, Grenouillet Frédéric, Quadroni Manfredo, Fellrath Jean-Marc, Aubert John-David, Dalphin Jean-Charles, Millon Laurence
UMR Chrono-Environnement 6249/CNRS, University of Besancon, Place Saint-Jacques, Besançon 25030 cedex, France.
Clin Vaccine Immunol. 2010 Jan;17(1):160-7. doi: 10.1128/CVI.00129-09. Epub 2009 Nov 11.
In France and Finland, farmer's lung disease (FLD), a hypersensitivity pneumonitis common in agricultural areas, is mainly caused by Eurotium species. The presence of antibodies in patients' serum is an important criterion for diagnosis. Our study aimed to improve the serological diagnosis of FLD by using common fungal particles that pollute the farm environment as antigens. Fungal particles of the Eurotium species were observed in handled hay. A strain of Eurotium amstelodami was grown in vitro using selected culture media; and antigen extracts from sexual (ascospores), asexual (conidia), and vegetative (hyphae) forms were made. Antigens were tested by enzyme-linked immunosorbent assay (ELISA), which was used to test for immunoglobulin G antibodies from the sera of 17 FLD patients, 40 healthy exposed farmers, and 20 nonexposed controls. The antigens were compared by receiver operating characteristic analysis, and a threshold was then established. The ascospores contained in asci enclosed within cleistothecia were present in 38% of the hay blades observed; conidial heads of aspergillus were less prevalent. The same protocol was followed to make the three antigen extracts. A comparison of the results for FLD patients and exposed controls showed the area under the curve to be 0.850 for the ascospore antigen, 0.731 for the conidia, and 0.690 for the hyphae. The cutoffs that we determined, with the standard deviation for measures being taken into account, showed 67% for sensitivity and 92% for specificity with the ascospore antigen. In conclusion, the serological diagnosis of FLD by ELISA was improved by the adjunction of ascospore antigen.
在法国和芬兰,农民肺疾病(FLD)是农业地区常见的一种超敏性肺炎,主要由曲霉菌属引起。患者血清中抗体的存在是诊断的重要标准。我们的研究旨在通过使用污染农场环境的常见真菌颗粒作为抗原,改进FLD的血清学诊断。在处理过的干草中观察到曲霉菌属的真菌颗粒。使用选定的培养基在体外培养一株阿姆斯特丹曲霉;并制备了来自有性(子囊孢子)、无性(分生孢子)和营养(菌丝)形式的抗原提取物。通过酶联免疫吸附测定(ELISA)对抗原进行检测,该方法用于检测17例FLD患者、40名健康暴露农民和20名非暴露对照者血清中的免疫球蛋白G抗体。通过受试者工作特征分析对抗原进行比较,然后确定一个阈值。在观察到的38%的干草叶片中,闭囊壳内包含的子囊中的子囊孢子存在;曲霉的分生孢子头则不太常见。制备三种抗原提取物遵循相同的方案。对FLD患者和暴露对照者的结果比较显示,子囊孢子抗原的曲线下面积为