时间分辨荧光免疫分析高灵敏检测嗜水气单胞菌
发布时间：2018-07-23 12:02
【摘要】：建立了生物素-链霉亲和素时间分辨荧光免疫分析(BAS-TRFIA)高灵敏检测嗜水气单胞菌的新方法.以兔抗IgG包被微孔板,加入嗜水气单胞菌菌株B18和生物素化IgG,生成的夹心复合物用Eu~(3+)-链霉亲和素作为示踪物,Eu~(3+)可与离解增强液形成时间分辨荧光(TRF),通过TRF值对病原菌定量.方法的检测限为1.0×10~2 cfu/mL,在1.0×10~1.0×10~6 cfu/mL范围内线性良好,相关系数0.9716.用该法检测时,不同来源的嗜水气单胞菌以及其它气单胞菌均呈阴性.板内和板间的变异系数分别小于5.00%和9.00%.所有相关检测试剂37℃恒温放置6 d后,检测性能无明显改变.对60份人工感染的美洲鳗鲡组织样品,包括肝、肾、肠、鳃和肌肉等组织进行检测,阳性检测率为98.33%.结果表明,BAS-TRFIA法检测嗜水气单胞菌,灵敏度高、特异性好、操作简便,该方法为水产养殖病原菌检测提供了新的思路.
[Abstract]:A highly sensitive method for the detection of Aeromonas hydrophila by biotin-streptavidin time resolved fluorescence immunoassay (BAS-TRFIA) was established. The micropore plate of rabbit anti IgG was coated with B18 strain of Aeromonas hydrophila and biotinylated IgG. The sandwich complex was produced with EU3-Streptomyces avidin as a tracer, which could be used to form a time resolved fluorescent (TRF), with dissociation enhancer to quantify the pathogenic bacteria by TRF value. The detection limit of the method is 1.0 脳 10 ~ (-2) cfur 路mL ~ (-1), the linear range is 1.0 脳 10 ~ (10) 脳 10 ~ (-6) cfu/mL, the correlation coefficient is 0.9716. When detected by this method, Aeromonas hydrophila and other Aeromonas from different sources were negative. The coefficient of variation within and between boards was less than 5.00% and 9.00%, respectively. The detection performance of all the related reagents remained unchanged after 6 days at 37 鈩，
本文编号：2139350
[Abstract]:A highly sensitive method for the detection of Aeromonas hydrophila by biotin-streptavidin time resolved fluorescence immunoassay (BAS-TRFIA) was established. The micropore plate of rabbit anti IgG was coated with B18 strain of Aeromonas hydrophila and biotinylated IgG. The sandwich complex was produced with EU3-Streptomyces avidin as a tracer, which could be used to form a time resolved fluorescent (TRF), with dissociation enhancer to quantify the pathogenic bacteria by TRF value. The detection limit of the method is 1.0 脳 10 ~ (-2) cfur 路mL ~ (-1), the linear range is 1.0 脳 10 ~ (10) 脳 10 ~ (-6) cfu/mL, the correlation coefficient is 0.9716. When detected by this method, Aeromonas hydrophila and other Aeromonas from different sources were negative. The coefficient of variation within and between boards was less than 5.00% and 9.00%, respectively. The detection performance of all the related reagents remained unchanged after 6 days at 37 鈩，
本文编号：2139350
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