一种沙门氏菌化学发光酶免疫分析方法的建立  

Establishment of chemiluminescent enzyme immunoassay for Salmonella

作　　者：房芳[1,2] 敬思群[1] 李锦丰[2] 曹际娟[3] 徐静[3] 曲圣轩 金福姝[2] 邹明强[2] 

机构地区：[1]新疆大学生命科学与技术学院,乌鲁木齐830046 [2]中国检验检疫科学研究院,北京100123 [3]辽宁出入境检验检疫局检验检疫技术中心,大连116001 [4]大连医诺生物有限公司,大连116600

出　　处：《食品科技》2014年第8期281-286,共6页Food Science and Technology

基　　金："十二五"国家科技支撑计划项目(2011BAK10B02)

摘　　要：主要研究沙门氏菌的化学发光酶免疫(CLEIA)快速检测方法。以猪霍乱沙门氏菌(Salmonella choleraesuis)、鼠伤寒沙门氏菌(Salmonella typhimurium)、肠炎沙门氏菌(Salmonella enteritidis)作为抗原,沙门氏菌多克隆抗体作为一抗,辣根过氧化物酶标记的山羊抗兔IgG(IgGHRP)作为酶标二抗,选用HRP-鲁米诺-H2O2化学发光体系,通过优化底物中各组分条件,建立了一种可同时检测3种沙门氏菌的间接CLEIA方法,此方法检测限为1×104 cfu/mL,且与其他3种常见肠道致病菌均无交叉反应。对人工添加牛乳样品预增菌后,应用建立的沙门氏菌间接CLEIA快速检测方法进行沙门氏菌检测,检测准确率可达到97.33%。该研究建立的沙门氏菌间接CLEIA快速检测方法简便、快速、准确,具有良好的特异性和重复性。A rapid and sensitive chemiluminescent enzyme immunoassay（CLEIA） method has been developed for the determination of the Salmonella. The method development was processed with salmonella polyclonal antibody and goat anti-rabbit IgG labeled by HRP, as well as, HRP-LuminolH2O2 chemiluminescent system. The method was developed for determinate Salmonella choleraesuis, Salmonella typhimurium and Salmonella enteritidis in milk at same time. The conditions has been optimized, the detection limit was 1×104 cfu/mL. Meanwhile, no cross-reaction with the other 3 strains ofintestinal bacteria was found. The milk sample which has artifical additive has been used for proliferation before determination. The accuracy of this method with Salmonella determination was 97.33%. A high throughput method for the determination of Salmonella by CLEIA has been developed due to its feature of rapid, sensitive, simple operation, good specificity and good reproducibility.

关 键 词：沙门氏菌 化学发光免疫酶检测 底物优化 

分 类 号：TS201.3[轻工技术与工程—食品科学]