van der Poel W H M, Pavio N, van der Goot J, van Es M, Martin M, Engel B
Central Veterinary Institute, Wageningen University and Research Centre, Lelystad, The Netherlands.
Animal Health Laboratory UMR 1161, Agence Nationale de Sécurité Sanitaire de l'Alimentation, de l'Environnement et du Travail, Maisons-Alfort, France.
Braz J Med Biol Res. 2014 Apr;47(4):334-9. doi: 10.1590/1414-431x20133249. Epub 2014 Mar 21.
Hepatitis E virus (HEV) is classified within the family Hepeviridae, genus Hepevirus. HEV genotype 3 (Gt3) infections are endemic in pigs in Western Europe and in North and South America and cause zoonotic infections in humans. Several serological assays to detect HEV antibodies in pigs have been developed, at first mainly based on HEV genotype 1 (Gt1) antigens. To develop a sensitive HEV Gt3 ELISA, a recombinant baculovirus expression product of HEV Gt3 open reading frame-2 was produced and coated onto polystyrene ELISA plates. After incubation of porcine sera, bound HEV antibodies were detected with anti-porcine anti-IgG and anti-IgM conjugates. For primary estimation of sensitivity and specificity of the assay, sets of sera were used from pigs experimentally infected with HEV Gt3. For further validation of the assay and to set the cutoff value, a batch of 1100 pig sera was used. All pig sera were tested using the developed HEV Gt3 assay and two other serologic assays based on HEV Gt1 antigens. Since there is no gold standard available for HEV antibody testing, further validation and a definite setting of the cutoff of the developed HEV Gt3 assay were performed using a statistical approach based on Bayes' theorem. The developed and validated HEV antibody assay showed effective detection of HEV-specific antibodies. This assay can contribute to an improved detection of HEV antibodies and enable more reliable estimates of the prevalence of HEV Gt3 in swine in different regions.
戊型肝炎病毒（HEV）属于戊型肝炎病毒科戊型肝炎病毒属。戊型肝炎病毒3型（Gt3）感染在西欧以及南北美洲的猪中呈地方性流行，并可导致人类发生人畜共患病感染。已经开发了几种用于检测猪体内戊型肝炎病毒抗体的血清学检测方法，最初主要基于戊型肝炎病毒1型（Gt1）抗原。为了开发一种灵敏的戊型肝炎病毒Gt3酶联免疫吸附测定（ELISA），制备了戊型肝炎病毒Gt3开放阅读框2的重组杆状病毒表达产物，并将其包被在聚苯乙烯ELISA板上。在孵育猪血清后，用抗猪抗IgG和抗IgM缀合物检测结合的戊型肝炎病毒抗体。为了初步评估该检测方法的敏感性和特异性，使用了经戊型肝炎病毒Gt3实验感染的猪的血清样本。为了进一步验证该检测方法并设定临界值，使用了一批1100份猪血清。所有猪血清均使用所开发的戊型肝炎病毒Gt3检测方法以及另外两种基于戊型肝炎病毒Gt1抗原的血清学检测方法进行检测。由于目前尚无戊型肝炎病毒抗体检测的金标准，因此基于贝叶斯定理采用统计方法对所开发的戊型肝炎病毒Gt3检测方法进行了进一步验证并确定了临界值。所开发并经验证的戊型肝炎病毒抗体检测方法能够有效检测戊型肝炎病毒特异性抗体。该检测方法有助于改进戊型肝炎病毒抗体的检测，并能更可靠地估计不同地区猪群中戊型肝炎病毒Gt3的流行率。
