Navarro Alejandro, Bárcena Carmen, Pozo Pilar, Díez-Guerrier Alberto, Martínez Irene, Polo Coral, Duque Clara, Rodríguez-Lázaro David, Goyache Joaquín, García Nerea
VISAVET Health Surveillance Center, Complutense University of Madrid, 28040 Madrid, Spain.
MAEVA SERVET, S.L., Alameda del Valle, 28749 Madrid, Spain.
Microorganisms. 2020 Mar 23;8(3):450. doi: 10.3390/microorganisms8030450.
In recent years, cases of hepatitis E virus (HEV) infection have increased in Europe in association with the consumption of contaminated food, mainly from pork products but also from wild boars. The animal's serum is usually tested for the presence of anti-HEV antibodies and viral RNA but, in many cases such as during hunting, an adequate serum sample cannot be obtained. In the present study, liver transudate was evaluated as an alternative matrix to serum for HEV detection. A total of 125 sera and liver transudates were tested by enzyme-linked immunosorbent assay at different dilutions (1:2, 1:10, 1:20), while 58 samples of serum and liver transudate were checked for the presence of HEV RNA by RT-qPCR. Anti- HEV antibodies were detected by ELISA in 68.0% of the serum samples, and in 61.6% of the undiluted transudate, and in 70.4%, 56.8%, and 44.8% of 1:2, 1:10, or 1:20 diluted transudate, respectively. The best results were obtained for the liver transudate at 1:10 dilution, based on the Kappa statistic (0.630) and intraclass correlation coefficient (0.841). HEV RNA was detected by RT-qPCR in 22.4% of the serum samples and 6.9% of the transudate samples, all samples used for RT-qPCR were positive by ELISA. Our results indicate that liver transudate may be an alternative matrix to serum for the detection of anti-HEV antibodies.
近年来,欧洲戊型肝炎病毒(HEV)感染病例有所增加,这与食用受污染食物有关,主要是猪肉制品,也包括野猪。通常会检测动物血清中抗HEV抗体和病毒RNA的存在,但在许多情况下,如狩猎期间,无法获得足够的血清样本。在本研究中,评估了肝渗出液作为血清的替代基质用于检测HEV。通过酶联免疫吸附测定法在不同稀释度(1:2、1:10、1:20)下对总共125份血清和肝渗出液进行了检测,同时通过逆转录定量聚合酶链反应(RT-qPCR)检查了58份血清和肝渗出液样本中HEV RNA的存在情况。通过酶联免疫吸附测定法在68.0%的血清样本、61.6%的未稀释渗出液以及分别在70.4%、56.8%和44.8%的1:2、1:10或1:20稀释渗出液中检测到抗HEV抗体。基于卡帕统计量(0.630)和组内相关系数(0.841),在1:10稀释的肝渗出液中获得了最佳结果。通过RT-qPCR在22.4%的血清样本和6.9%的渗出液样本中检测到HEV RNA,所有用于RT-qPCR的样本通过酶联免疫吸附测定法均呈阳性。我们的结果表明,肝渗出液可能是检测抗HEV抗体时血清的替代基质。