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用于检测巴西南部猪群中常见沙门氏菌血清型抗体的酶联免疫吸附测定法的开发与应用

Development and application of an enzyme-linked immunosorbent assay to detect antibodies against prevalent Salmonella serovars in swine in southern Brazil.

作者信息

Kich Jalusa Deon, Schwarz Patrícia, Eduardo Silva Luis, Coldebella Arlei, Piffer Itamar Antônio, Vizzoto Remídio, Ribeiro de Itapema Cardoso Marisa

机构信息

Embrapa Suínos e Aves, Concórdia, SC, Brazil.

出版信息

J Vet Diagn Invest. 2007 Sep;19(5):510-7. doi: 10.1177/104063870701900508.

Abstract

The implementation of Salmonella control programs in the pork production chain demands rapid and cost-effective methods to assess the prevalence of infection in pig herds. The objective of the present study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) based on S. Typhimurium lipopolysaccharides (LPS) to measure the prevalence of infection caused by Salmonella in swine herds. Coating antigen was produced by phenol extraction of S. Typhimurium culture. After standardization of ELISA test conditions, the assay was validated by testing serum samples on different animal categories: pigs orally inoculated with S. Typhimurium and sentinel animals in contact with them, naturally infected animals, colostrum-deprived piglets, and bacterin-immunized pigs. Seroconversion was observed in inoculated pigs (7 days postinfection [DPI]) and in the sentinels (21 DPI). Nonspecific reactions were not detected in the sera of colostrum-deprived animals. Serum samples from animals immunized with Salmonella Agona, Salmonella Derby, Salmonella Panama, and Salmonella Bredeney bacterins showed marked cross-reaction with the LPS from the serovar Typhimurium. Moreover, positive results obtained with the in-house ELISA were associated with Salmonella isolation in 75 infected pig herds. Comparisons with 2 commercial kits showed a linear correlation coefficient of 0.847 between the in-house ELISA and kit A and 0.922 with kit B but a low agreement in the qualitative results. In conclusion, the newly developed in-house ELISA based on S. Typhimurium LPS can be a useful tool to determine the intensity of Salmonella sp. infection in swine herds.

摘要

在猪肉生产链中实施沙门氏菌控制计划需要快速且经济高效的方法来评估猪群中的感染率。本研究的目的是开发一种基于鼠伤寒沙门氏菌脂多糖(LPS)的内部酶联免疫吸附测定(ELISA),以测量猪群中由沙门氏菌引起的感染率。包被抗原通过鼠伤寒沙门氏菌培养物的苯酚提取制备。在ELISA试验条件标准化后,通过检测不同动物类别的血清样本对该测定进行验证:口服接种鼠伤寒沙门氏菌的猪及其接触的哨兵动物、自然感染动物、初乳剥夺仔猪和菌苗免疫猪。在接种猪(感染后7天[DPI])和哨兵动物(21 DPI)中观察到血清转化。在初乳剥夺动物的血清中未检测到非特异性反应。用阿贡纳沙门氏菌、德比沙门氏菌、巴拿马沙门氏菌和布雷登尼沙门氏菌菌苗免疫的动物的血清样本与鼠伤寒血清型的LPS显示出明显的交叉反应。此外,内部ELISA获得的阳性结果与75个感染猪群中的沙门氏菌分离相关。与2种商业试剂盒的比较显示,内部ELISA与试剂盒A之间的线性相关系数为0.847,与试剂盒B之间为0.922,但定性结果的一致性较低。总之,新开发的基于鼠伤寒沙门氏菌LPS的内部ELISA可作为确定猪群中沙门氏菌感染强度的有用工具。

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