旋毛虫--使用化学发光免疫测定法制备样品和客观检测特异性抗原的新方法。

Q1 Veterinary Veterinary Parasitology: X Pub Date : 2020-10-08 eCollection Date: 2020-12-01 DOI:10.1016/j.vpoa.2020.100033
Jana Braasch, Stefanie Ostermann, Monika Mackiewicz, Catherine Bardot, Caroline Pagneux, Viola Borchardt-Lohölter, Erik Lattwein
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引用次数: 0

摘要

螺旋毛蛔虫最常见的传播途径是食用生的或未煮熟的受感染猪肉或野味。为防止人类感染,屠宰场使用黄金标准 "磁力搅拌器法 "进行肉类安全监测。我们采用新开发的基于化学发光的免疫测定法(ChLIA)自动检测特异性旋毛虫抗原,从而推出了一种快速、客观的方法。样本 A 包括未感染猪的肌肉组织样本(n = 37)。B 组为未感染猪的肌肉组织样本,其中添加了不同数量的毛滴虫幼虫,但不含胶原蛋白胶囊(n = 56)。C 组为实验感染猪的肌肉组织样本,包括胶原蛋白包裹的毛滴虫幼虫(n = 32)。每个样本都在刀磨机中用 PBS 缓冲液切碎,以消灭毛线虫幼虫。离心后,上清液(肌肉组织提取物,其中含有释放的排泄和分泌螺旋体旋毛虫抗原)用于新型旋毛虫 ChLIA 检测旋毛虫特异性抗原。ChLIA 的总体准确率为 97.6%。ChLIA 的特异性为 100%(A 组)。实验感染猪样本的灵敏度为 100%,检测限为每克 0.01 个幼虫。未观察到与旋毛虫属以外的寄生虫的交叉反应。这种用于检测旋毛虫抗原的新型肉类检测方法具有高特异性和高灵敏度,特别是在真正感染的样本中。与黄金标准相比,这种新的肉类安全监测方法不需要训练有素的人员进行长时间的消化或显微镜检查。
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Trichinella spiralis - New method for sample preparation and objective detection of specific antigens using a chemiluminescence immunoassay.

The parasitic roundworm Trichinella spiralis is most commonly transmitted to humans through consumption of raw or undercooked meat of infected pigs or game. To prevent human infection, slaughterhouses perform meat safety surveillance using the gold standard "Magnetic Stirrer Method". We introduce a fast and objective method using automated detection of specific Trichinella spiralis antigens by a newly developed immunoassay based on chemiluminescence (ChLIA). Panel A comprised muscle tissue samples from non-infected pigs (n = 37). Panel B comprised muscle tissue samples from non-infected pigs spiked with different amounts of Trichinella larvae without collagen capsules (n = 56). Panel C contained muscle tissue samples from experimentally infected pigs including Trichinella larvae encapsulated in collagen (n = 32). Each sample was shredded with PBS buffer in a knife mill, destroying Trichinella larvae. Following centrifugation, the supernatant (muscle tissue extract containing released excretory and secretory Trichinella spiralis antigens) was used for Trichinella-specific antigen detection by the new Trichinella ChLIA. The overall accuracy of the Trichinella ChLIA was 97.6 %. The specificity of the Trichinella ChLIA was 100 % (panel A). The sensitivity in samples from experimentally infected pigs was 100 % representing a detection limit of 0.01 larvae per gram. Cross-reactivity with parasites other than Trichinella spp. was not observed. This new meat inspection method for the detection of Trichinella spiralis antigens presents high specificity and high sensitivity, especially in truly infected samples. In contrast to the gold standard, this new approach to meat safety surveillance does not require longsome digestion or microscopy by trained personnel.

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Veterinary Parasitology: X
Veterinary Parasitology: X Veterinary-Veterinary (all)
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