Flannery John, Moore Rebecca, Marsella Laura, Harris Katie, Ashby Martin, Rajko-Nenow Paulina, Roberts Helen, Gubbins Simon, Batten Carrie
The Pirbright Institute, Ash Road, Pirbright GU24 0NF, UK.
Defra, Nobel House, 17 Smith Square, London SW1P 3JR, UK.
Foods. 2020 Aug 20;9(9):1148. doi: 10.3390/foods9091148.
African swine fever (ASF) is a highly lethal disease of pigs caused by the ASF virus (ASFV), which presents a serious threat to global food security. The movement of contaminated pork products has previously been postulated as contributing to the introduction of ASF into new areas. To evaluate the performance of ASFV detection systems in multi-component pork products, we spiked sausage meat with four different ASFV-containing materials (ASFV cell culture, pork loin, meat juice and bone marrow). DNA was extracted using two manual systems (MagMAX CORE, Qiagen) and one automated (MagMAX CORE) one, and three qPCR assays (VetMAX, King, UPL) were used. The performance of the DNA extraction systems was as follows; automated MagMAX > manual MagMAX > manual Qiagen. The commercial VetMAX qPCR assay yielded significantly lower C values ( < 0.001), showing greater sensitivity than the World Organization for Animal Health (OIE)-prescribed assays (King, UPL). Detection probability was the highest for matrices contaminated with bone marrow compared with pork loin or meat juice. An estimated minimum sample size of one 1-g sample is sufficient to detect ASFV in a homogenous pork product if bone marrow from infected pigs comprises 1 part in 10,000. We demonstrated that existing ASFV detection systems are appropriate for use in a food-testing capacity, which can provide an additional control measure for ASF.
非洲猪瘟（ASF）是由非洲猪瘟病毒（ASFV）引起的一种猪的高致死性疾病，对全球粮食安全构成严重威胁。此前曾推测受污染猪肉产品的流通是导致ASF传入新地区的原因之一。为评估ASFV检测系统在多成分猪肉产品中的性能，我们用四种不同的含ASFV材料（ASFV细胞培养物、猪里脊肉、肉汁和骨髓）对香肠肉进行了加标。使用两种手动系统（MagMAX CORE、Qiagen）和一种自动化系统（MagMAX CORE）提取DNA，并使用三种qPCR检测方法（VetMAX、King、UPL）。DNA提取系统的性能如下：自动化的MagMAX＞手动的MagMAX＞手动的Qiagen。商业VetMAX qPCR检测方法产生的C值显著更低（＜0.001），显示出比世界动物卫生组织（OIE）规定的检测方法（King、UPL）更高的灵敏度。与猪里脊肉或肉汁相比，受骨髓污染的基质的检测概率最高。如果感染猪的骨髓在均质猪肉产品中占万分之一，估计一个1克样品的最小样本量就足以检测到ASFV。我们证明现有的ASFV检测系统适用于食品检测，可为非洲猪瘟提供额外的控制措施。
