基于BHHCT-Eu3+@SiO2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

采用微波辅助合成的荧光稀土二氧化硅纳米颗粒(BHHCT-Eu3+@SiO2)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-Eu3+@SiO2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-Eu3+@SiO2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Eu3+-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(IC10)为0.85 ng/mL,半数抑制浓度(IC50)为12.76 ng/mL,检测范围(IC20-IC80)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.

Development of Immunochromatographic Strips Based on Covalently Conjugated BHHCT-Eu3+@SiO2 for Rapid and Quantitative Detection of Kanamycin

As a kind of fluorescent nanoprobe, BHHCT-Eu3+ @ SiO2 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-Eu3+@SiO2 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-Eu3+@SiO2 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 IC50 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 was<1%. The detection results of kana in milk samples between the LFIA and traditional ELISA method showed good correlation.