Brigmon R L, Zam S G, Wilson H R
Department of Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville 32611, USA.
Poult Sci. 1995 Jul;74(7):1232-6. doi: 10.3382/ps.0741232.
An ELISA previously developed for the rapid detection of Salmonella enteritidis (SE) in environmental samples was modified and applied to food samples. A sandwich ELISA was designed that employs affinity-purified rabbit polyclonal antibodies for the capture stage and highly specific monoclonal antibodies for the detection stage. Thirty-nine species of bacteria other than SE, including 32 Salmonella species, were included in cross-reactivity testing with ELISA. Results showed no reactivity with any species tested besides SE. Salmonella enteritidis was added to homogenized food samples (chicken skin, meat, and eggs) to test ELISA sensitivity. The lower limit for ELISA detection of SE was 10(4) cells/mL for pure cultures and in 10% meat (wt/vol), 10(5) cells/mL in 10% skin (wt/vol), and 10(7) cells/mL in 10% eggs (wt/vol). Salmonella enteritidis detection with ELISA was confirmed with the Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) method. Results were obtained within 24 h for ELISA method compared to 96 h for the BAM procedure. Results show that sensitivity of ELISA can vary with the type of food tested for detection of SE.
一种先前开发用于快速检测环境样品中肠炎沙门氏菌（SE）的酶联免疫吸附测定（ELISA）方法经过改良后应用于食品样品。设计了一种夹心ELISA，在捕获阶段采用亲和纯化的兔多克隆抗体，在检测阶段采用高特异性单克隆抗体。除SE外的39种细菌，包括32种沙门氏菌，都参与了ELISA的交叉反应性测试。结果表明，除SE外，与所测试的任何物种均无反应性。将肠炎沙门氏菌添加到匀浆食品样品（鸡皮、肉和蛋）中以测试ELISA的灵敏度。对于纯培养物以及在10%的肉（重量/体积）中，ELISA检测SE的下限为10⁴个细胞/毫升，在10%的皮（重量/体积）中为10⁵个细胞/毫升，在10%的蛋（重量/体积）中为10⁷个细胞/毫升。ELISA检测肠炎沙门氏菌的结果通过美国食品药品监督管理局（FDA）的《细菌学分析手册》（BAM）方法得到了确认。ELISA方法在24小时内获得结果，而BAM程序则需要96小时。结果表明，ELISA的灵敏度会因所检测食品的类型而异，用于检测SE时情况也是如此。
