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评价 Western Blot 和 ELISA 检测猪血清中抗旋毛虫 IgG 的效果。

Evaluation of a Western Blot and ELISA for the detection of anti-Trichinella-IgG in pig sera.

机构信息

Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany.

出版信息

Vet Parasitol. 2009 Aug 26;163(4):341-7. doi: 10.1016/j.vetpar.2009.04.034. Epub 2009 May 4.

Abstract

Human trichinellosis is a foodborne disease caused by ingestion of infective Trichinella muscle larvae via pork or meat of other food animals which are susceptible to this zoonotic parasite. There are new approaches for a risk-oriented meat inspection for Trichinella in pigs which are accompanied by monitoring programmes on herd level to control freedom from this parasite. For this purpose, testing schemes utilizing serological tests with a high sensitivity and specificity are required. This study aimed at the evaluation of an ELISA and a Western Blot (WB) for the detection of anti-Trichinella-IgG in terms of sensitivity and specificity taking results of artificial digestion as gold standard. For this purpose, 144 field sera from pigs confirmed as Trichinella-free as well as 159 sera from pigs experimentally infected with T. spiralis (123), T. britovi (19) or T. pseudospiralis (17) were examined by ELISA (excretory-secretory antigen) and WB (crude worm extract). Sera from pigs experimentally infected with four other nematode species were included to investigate the cross-reactivity of the antigen used in the WB. For all Trichinella-positive pig sera, band pattern profiles were identified in the WB and results were analysed in relation to ELISA OD% values. Testing of pig sera revealed a sensitivity of 96.8% for the ELISA and 98.1% for the WB whereas the methods showed a specificity of 97.9 and 100%, respectively. WB analysis of Trichinella-positive pig sera revealed five specific band patterns of 43, 47, 61, 66, and 102 kDa of which the 43 kDa protein was identified as the predominant antigen. The frequency of the band pattern profile was irrespective of the dose and the period of infection as well as the Trichinella species investigated. In conclusion, monitoring in swine farms for Trichinella antibodies should be based on screening pig sera by means of ELISA followed by confirmatory testing through WB analysis.

摘要

人旋毛虫病是一种食源性疾病,由摄入感染性旋毛虫肌肉幼虫引起,这些幼虫通过猪肉或其他易受这种人畜共患寄生虫感染的食用动物传播。目前有新的方法可以对猪进行针对旋毛虫的风险导向性肉检,并通过畜群监测计划来控制这种寄生虫的感染。为此,需要利用具有高灵敏度和特异性的血清学检测方法。本研究旨在评估 ELISA 和 Western Blot(WB)检测抗旋毛虫 IgG 的敏感性和特异性,以人工消化结果为金标准。为此,检测了 144 份来自经证实无旋毛虫感染的猪的田间血清样本,以及 159 份来自经实验感染旋毛虫(123 份)、旋毛虫(19 份)或旋毛虫(17 份)的猪的血清样本。ELISA(排泄分泌抗原)和 WB(粗虫体提取物)。还包括来自实验感染另外四种线虫物种的猪血清,以研究 WB 中使用的抗原的交叉反应性。对于所有旋毛虫阳性猪血清,WB 中均鉴定出了带型谱,分析结果与 ELISA OD%值相关。猪血清检测显示,ELISA 的敏感性为 96.8%,WB 的敏感性为 98.1%,而这两种方法的特异性分别为 97.9%和 100%。WB 分析旋毛虫阳性猪血清显示,有 5 种特异性带型谱,分子量分别为 43、47、61、66 和 102 kDa,其中 43 kDa 蛋白被鉴定为主要抗原。带型谱的频率与剂量、感染期以及所研究的旋毛虫种类无关。总之,对养猪场的旋毛虫抗体进行监测,应基于 ELISA 筛选猪血清,然后通过 WB 分析进行确认检测。

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