Department of Virology, National Reference Center for HEV, CHU Purpan, 31059 Toulouse, France.
Centre de Physiopathologie de Toulouse Purpan (CPTP), Institut National de la Santé et de la Recherche médicale, Inserm UMR1043, Centre national de la recherche scientifique, CNRS UMR5282, Université de Toulouse, 31024 Toulouse, France.
Viruses. 2020 Jan 24;12(2):139. doi: 10.3390/v12020139.
Hepatitis E virus (HEV) is a major concern in public health worldwide. Infections with HEV genotypes 3, 4, or 7 can lead to chronic hepatitis while genotype 1 infections can trigger severe hepatitis in pregnant women. Infections with all genotypes can worsen chronic liver diseases. As virions are lipid-associated in blood and naked in feces, efficient methods of propagating HEV clinical strains in vitro and evaluating the infectivity of both HEV forms are needed. We evaluated the spread of clinical strains of HEV genotypes 1 (HEV1) and 3 (HEV3) by quantifying viral RNA in culture supernatants and cell lysates. Infectivity was determined by endpoint dilution and calculation of the tissue culture infectious dose 50 (TCID50). An enhanced HEV production could be obtained varying the composition of the medium, including fetal bovine serum (FBS) and dimethylsulfoxide (DMSO) content. This increased TCID50 from 10 to 100-fold and allowed us to quantify HEV1 infectivity. These optimized methods for propagating and measuring HEV infectivity could be applied to health safety processes and will be useful for testing new antiviral drugs.
戊型肝炎病毒（HEV）是全球公共卫生的主要关注点。感染 HEV 基因型 3、4 或 7 可导致慢性肝炎，而基因型 1 感染可在孕妇中引发严重肝炎。所有基因型的感染均可使慢性肝病恶化。由于病毒粒子在血液中与脂质相关，而在粪便中呈裸露状态，因此需要有效的方法在体外繁殖 HEV 临床株并评估两种 HEV 形式的感染性。我们通过定量检测培养上清液和细胞裂解物中的病毒 RNA 来评估 HEV 基因型 1（HEV1）和 3（HEV3）临床株的传播。通过终点稀释和计算组织培养感染剂量 50（TCID50）来确定感染性。通过改变培养基的组成，包括胎牛血清（FBS）和二甲基亚砜（DMSO）的含量，可以获得增强的 HEV 产生。这将 TCID50 提高了 10 到 100 倍，并使我们能够定量检测 HEV1 的感染性。这些用于繁殖和测量 HEV 感染性的优化方法可应用于健康安全过程，并将有助于测试新的抗病毒药物。
