基于BHHCT-Eu^(3+)@SiO_2荧光稀土二氧化硅纳米颗粒的免疫层析试纸条检测卡那霉素

Development of Immunochromatographic Strips Based on Covalently Conjugated BHHCT-Eu^(3+)@SiO_2 for Rapid and Quantitative Detection of Kanamycin

采用微波辅助合成的荧光稀土二氧化硅纳米颗粒(BHHCT-Eu^(3+)@SiO_2)为标记物,建立了快速定量检测卡那霉素(Kana)残留的荧光免疫层析方法。实验结果表明,微波辅助合成的BHHCT-Eu^(3+)@SiO_2纳米颗粒呈球形,粒径约36 nm,具有良好的荧光发射性能,最大吸收波长和最大发射波长分别为343和615 nm。将BHHCT-Eu^(3+)@SiO_2与卡那霉素抗体(Kana-ab)通过醛基化葡聚糖交联,合成了荧光标记抗体Eu^(3+)-Kana-ab,结合定量侧向层析读数仪,建立了牛奶中Kana残留的快速定量检测方法,对Kana的检出限(IC_(10))为0.85 ng/mL,半数抑制浓度(IC_(50))为12.76 ng/mL,检测范围(IC_(20)-IC_(80))为3.0~76.0 ng/mL,牛奶中的Kana的加标回收率范围为93.7%~97.4%,RSD为3.1%~4.6%,与Kana类似物的交叉反应均<1%。牛奶中Kana残留的测定结果与ELISA方法相关性良好。

As a kind of fluorescent nanoprobe,BHHCT-Eu^3＋@SiO_2 fluorescent nanoparticles were synthesized using microwave irradiation.Transmission electron microscopy characterization showed that the nanoparticles were in spherical shape with particle size of about 36 nm.The BHHCT-Eu^3＋@SiO_2 exhibited good fluorescence property,with a maximal excitation wavelength of 343 nm and an maximal emission wavelength of 615 nm.The fluorescence lateral flow immunoassay（LFIA） was established for rapid and quantitative detection of kanamycin（Kana） in milk after BHHCT-Eu^3＋@SiO_2 fluorescent nanoparticles were conjugated with Kana antibody,with dextran as a linker.The limit of detection of Kana with the LFIA method was 0.85 ng/mL with IC_（50） of 12.76 ng/mL,and the detection range was from 3.0 ng/m to 76 ng/mL.The recoveries of Kana in milk were between 93.7% and 97.4% with RSD of 3.1%-4.6%,and cross-reactivity of the fluorescence immunoassay strip for Kana determination was1%.The detection results of kana in milk samples between the LFIA and traditional ELISA method showed good correlation.