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提取用于酶免疫测定的不可沉淀形式的沙门氏菌抗原。

Extraction of Salmonella antigens in non-sedimentable forms for enzyme immunoassay.

作者信息

Blais B W, Yamazaki H

机构信息

Department of Biology, Carleton University, Ottawa, Ontario, Canada.

出版信息

Int J Food Microbiol. 1989 Aug;9(1):63-71. doi: 10.1016/0168-1605(89)90038-x.

Abstract

Heating Salmonella typhimurium in ethylenediaminetetraacetate was found to dissociate its antigens into forms that are non-sedimentable at 10,000 x g. The treatment caused a marked increase in the rate of immunoreaction and the sensitivity in an enzyme immunoassay for the detection of Salmonella antigens. The method permits the extraction of Salmonella antigens from solid-rich samples and the preparation of solid-free samples by means of centrifugation. When the level of the antigens in the supernatant was too low for the immunoassay, the antigens were readily concentrated by passing a large volume of the supernatant through a macroporous hydrophobic cloth coated with anti-Salmonella antibody. Using this method, the detection of as few as 10 Salmonella cells per gram of chicken meat was possible within a total of 18 h, which included 16 h of enrichment in either tetrathionate, selenite cystine, or nutrient broths.

摘要

研究发现,在乙二胺四乙酸中加热鼠伤寒沙门氏菌会使其抗原解离成在10,000×g离心力下不可沉降的形式。该处理显著提高了免疫反应速率以及用于检测沙门氏菌抗原的酶免疫测定的灵敏度。该方法允许从富含固体的样品中提取沙门氏菌抗原,并通过离心制备无固体样品。当上清液中抗原水平过低而无法进行免疫测定时,通过使大量上清液通过涂有抗沙门氏菌抗体的大孔疏水布,可轻松浓缩抗原。使用该方法,在总共18小时内,包括在四硫磺酸盐、亚硒酸盐胱氨酸或营养肉汤中富集16小时,每克鸡肉中低至10个沙门氏菌细胞也可被检测到。

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