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检测中国河北省生猪肉和猪内脏中的戊型肝炎病毒。

Detection of Hepatitis E Virus in Raw Pork and Pig Viscera As Food in Hebei Province of China.

机构信息

1 Health Science Center, Hebei University, Baoding, China.

2 Division of HIV/AIDS and Sex-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China.

出版信息

Foodborne Pathog Dis. 2019 May;16(5):325-330. doi: 10.1089/fpd.2018.2572. Epub 2019 Feb 26.

Abstract

Hepatitis E virus (HEV) is an important human pathogen with pigs serving as the main natural animal reservoir. In China, pork is the most popular meat, while pig viscera are also widely consumed. The aim of this study was to evaluate the prevalence of HEV among pigs at slaughter, and assess the presence of HEV in raw pork and pig viscera as food. Samples of pig blood, raw pork, liver, kidney, and blood curd were collected from slaughterhouse or (and) retail market. Anti-HEV antibody in serum samples was detected using enzyme-linked immunosorbent assay based on an ORF2 antigen sandwich kit. HEV RNA was tested by reverse transcription nested polymerase chain reaction (RT-nested PCR) and the viral load was further assessed using quantitative real-time PCR. The final amplicons of RT-nested PCR were sequenced and undergone phylogenetic analysis. Prevalence of antibodies to HEV was 90.4% (104/115) in pigs at slaughtered level, and one serum sample was HEV RNA positive (0.9%, 1/115). HEV RNA was detected in liver, kidney, and blood samples with positivity of 6.1% (7/114), 3.1% (4/129), and 1.2% (2/170) respectively with viral loads ranged 10-10 (2.4Log-4.4Log) genome equivalents per gram, but not in pork. The HEV RNA prevalence in both liver and kidney were statistically higher than in pork. Phylogenetic analysis showed that all obtained sequences belonged to HEV genotype 4, which were divided into subtypes 4a, 4b, 4d, and 4i, highly identical to the known human and swine HEV sequences in China. The results indicate that raw pig viscera are more likely to harbor HEV than pork, suggesting a higher transmission risk related to consuming pig organs.

摘要

戊型肝炎病毒(HEV)是一种重要的人类病原体,猪是其主要的天然动物宿主。在中国,猪肉是最受欢迎的肉类,而猪内脏也被广泛食用。本研究旨在评估屠宰猪中 HEV 的流行情况,并评估生猪肉和猪内脏作为食品时 HEV 的存在情况。采集自屠宰场或(和)零售市场的猪血液、生猪肉、肝脏、肾脏和血豆腐样本。采用基于 ORF2 抗原夹心试剂盒的酶联免疫吸附试验检测血清样本中的抗-HEV 抗体。采用反转录巢式聚合酶链反应(RT-nested PCR)检测 HEV RNA,并进一步采用实时荧光定量 PCR 评估病毒载量。对 RT-nested PCR 的最终扩增子进行测序并进行系统进化分析。屠宰猪中抗-HEV 抗体的流行率为 90.4%(104/115),1 份血清样本为 HEV RNA 阳性(0.9%,1/115)。在肝脏、肾脏和血样中检测到 HEV RNA,阳性率分别为 6.1%(7/114)、3.1%(4/129)和 1.2%(2/170),病毒载量范围为 10-10(2.4Log-4.4Log)基因组等价物/克,但未在猪肉中检测到。肝脏和肾脏中的 HEV RNA 流行率均明显高于猪肉。系统进化分析显示,所有获得的序列均属于 HEV 基因型 4,分为 4a、4b、4d 和 4i 亚型,与中国已知的人类和猪 HEV 序列高度同源。结果表明,生猪内脏比猪肉更有可能携带 HEV,提示与食用猪内脏相关的传播风险更高。

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