An ultrasensitive NIR-IIa’ fluorescence-based multiplex immunochromatographic strip test platform for antibiotic residues detection in milk samples,Journal of Advanced Research

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An ultrasensitive NIR-IIa’ fluorescence-based multiplex immunochromatographic strip test platform for antibiotic residues detection in milk samples
Journal of Advanced Research ( IF 11.4 ) Pub Date : 2022-10-22 , DOI: 10.1016/j.jare.2022.10.008
Yunyue Zhang ₁ , Tao Liao ₂ , Guoxin Wang ₂ , Juan Xu ₁ , Mohan Wang ₁ , Fazheng Ren ₃ , Hao Zhang ₃

Affiliation

Introduction

Widely used in livestock breeding, residues of antibiotic drugs in milk have become a threat to food safety and human health. Current rapid detection technologies using colorimetric immunochromatographic strip tests (IST) lack the necessary sensitivity for on-site trace monitoring. Fluorescence-based detection in the near-infrared IIa’ (NIR-IIa’) region (1000 ∼ 1300 nm) has enormous potential due to greatly minimized auto-fluorescence and light scattering.

Objectives

The aim of this work is to develop an ultrasensitive IST platform using NIR-IIa’ fluorescent nanoparticles as labels for multiplex antibiotic residues detection in milk.

Methods

NIR-IIa’ fluorescent nanoparticles were assembled by encapsulating synthesized NIR-IIa’ fluorophores into carboxyl - modified polystyrene nanoparticles. The NIR-IIa’ nanoparticles were subsequently used as labels in an IST platform to detect sulfonamides, quinolones, and lincomycin simultaneously in milk. A portable fluorescent reader was fabricated to provide on-site detection. To further validate the developed IST platform, the detection was compared with LC-MS/MS in 22 real milk samples.

Results

Fluorescent nanoparticles were synthesized with low energy emission (1030 nm) and large Stokes shift (>250 nm) showing a much higher signal-to-noise ratio compared with fluorophores emitting in the NIR-I region. The developed IST platform yielded a highly sensitive, simultaneous quantification of sulfonamides, quinolones, and lincomycin in milk with detection limits of 46.7, 27.6 and 51.4 pg/mL, respectively, achieving a wide detection range (up to 50 ng/mL). The IST platform showed good accuracy, reproducibility, and specificity with the portable fluorescent reader which could rapidly quantify in 10 s. These results were better than reported immunochromatographic assays using fluorescent labels, and remarkably, showed a higher recognition ability than LC-MS/MS for real samples.

Conclusion

The utility of NIR-IIa’ fluorescence-based IST platform for the fast, sensitive, and accurate detection of antibiotics in milk was demonstrated, successfully verifying the potential of this platform in detecting trace materials in complex matrices.

中文翻译：

用于牛奶样品中抗生素残留检测的超灵敏 NIR-IIa' 荧光多重免疫层析试纸测试平台

介绍

牛奶中广泛使用的抗生素药物残留已成为食品安全和人类健康的威胁。目前使用比色免疫层析试纸条测试 (IST) 的快速检测技术缺乏现场痕量监测所需的灵敏度。由于自发荧光和光散射大大减少，近红外 IIa' (NIR-IIa') 区域（1000 ∼ 1300 nm）基于荧光的检测具有巨大的潜力。

目标

这项工作的目的是开发一个超灵敏的 IST 平台，使用 NIR-IIa 荧光纳米颗粒作为牛奶中多重抗生素残留检测的标签。

方法

NIR-IIa'荧光纳米颗粒是通过将合成的NIR-IIa'荧光团封装到羧基改性的聚苯乙烯纳米颗粒中来组装的。NIR-IIa' 纳米颗粒随后被用作 IST 平台中的标记，以同时检测牛奶中的磺胺类药物、喹诺酮类药物和林可霉素。制造了便携式荧光读数器以提供现场检测。为了进一步验证开发的 IST 平台，在 22 个真实牛奶样品中将检测结果与 LC-MS/MS 进行了比较。

结果

合成的荧光纳米颗粒具有低能量发射 (1030 nm) 和大斯托克斯位移 (>250 nm)，与 NIR-I 区域发射的荧光团相比，显示出更高的信噪比。开发的 IST 平台可对牛奶中的磺胺类药物、喹诺酮类药物和林可霉素进行高灵敏度同步定量，检测限分别为 46.7、27.6 和 51.4 pg/mL，实现了较宽的检测范围（高达 50 ng/mL）。IST 平台通过便携式荧光读数器显示出良好的准确性、重现性和特异性，可在 10 秒内快速定量。这些结果优于报道的使用荧光标记的免疫层析测定，并且显着地显示出比 LC-MS/MS 对实际样品更高的识别能力。