Pancreatin is a combination of enzymes, principally amylase, lipase, and protease, used in the treatment of pancreatic endocrine insufficiency in humans. Pancreatin manufactured from imported porcine pancreas carries the risk of hepatitis E virus (HEV) contamination. About 1 % of the starting material for pancreatin manufacture is invariably constituted of the small intestine, which is known to be a major extrahepatic site of HEV replication in pigs. The aim of this study was to evaluate a method to detect and quantify HEV in pancreatin of porcine origin. Because HEV cannot be easily grown by conventional cell culture, an approach based on an established quantitative RT-PCR (RT-qPCR) was selected. This entailed the use of a non-HEV internal control to monitor RNA extraction efficacy and the production of HEV synthetic RNA as a reference to account for the efficacy of reverse-transcription. The method was evaluated by experiments in which HEV (from naturally infected pigs) was spiked in both the starting material (i.e., porcine pancreas homogenate for industrial production) and in the pancreatin itself. A laboratory protocol matching the industrial production workflow was set up and RT-qPCR experiments were carried out to evaluate the method's ability to detect HEV in pancreatin made from HEV-contaminated porcine tissues. The results showed that the method may be employed in two different strategies: to test the porcine pancreas homogenate (quantitative performance) or directly on pancreatin (qualitative assay). While the risk of HEV contamination in pancreatin may be low, it cannot be completely ruled out. Testing for HEV based on the precautionary principle ought to be the guiding rule.

中文翻译：

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评价猪源胰酶中戊型肝炎病毒（HEV）的分子方法。

胰酶是酶（主要是淀粉酶，脂肪酶和蛋白酶）的组合，用于治疗人的胰腺内分泌功能不全。由进口猪胰腺制造的胰酶具有戊型肝炎病毒（HEV）污染的风险。胰酶生产原料的大约1％总是由小肠组成，而小肠是猪中HEV复制的主要肝外部位。这项研究的目的是评估一种检测和定量猪源胰酶中HEV的方法。由于传统的细胞培养不能轻易培养HEV，因此选择了基于已建立的定量RT-PCR（RT-qPCR）的方法。这需要使用非HEV内部对照来监测RNA提取功效，并以HEV合成RNA的产生作为参考来解释逆转录功效。通过实验评估该方法，在该方法中，将HEV（来自自然感染的猪）加标在起始原料（即用于工业生产的猪胰腺匀浆）和胰酶本身中。建立了与工业生产流程匹配的实验室规程，并进行了RT-qPCR实验，以评估该方法检测由HEV污染的猪组织制成的胰酶中HEV的能力。结果表明，该方法可用于两种不同的策略：测试猪胰腺匀浆（定量性能）或直接在胰酶上（定性测定）。尽管胰酶中戊型肝炎病毒污染的风险可能较低，但不能完全排除它。基于预防原则的HEV测试应成为指导原则。