RESUMEN
The aim of this research was to develop a simple, rapid, sensitive, high-throughput detection method for foodborne Escherichia coli (E. coli) O157:H7 based on the aptamer-modified gold nanoparticles@macroporous magnetic silica photonic microsphere (Au@MMSPM). Such Au@MMSPM array system for E. coli O157:H7 not only integrated sample pretreatment with rapid detection, but also showed highly enhanced effect to develop a highly sensitive SERS assay. The established SERS assay platform gave a wide linear detection range (10-106 CFU/mL) and low limit of detection (2.20 CFU/mL) for E. coli O157:H7. The whole analysis time including sample pretreatment and detection was 110 min. This SERS-based assay platform provided a new high-throughput, highly sensitive and fast detection technology for monitoring E. coli O157:H7 in real samples from the fields of food industry, medicine and environment.
Asunto(s)
Técnicas Biosensibles , Escherichia coli O157 , Nanopartículas del Metal , Dióxido de Silicio , Oro , Microesferas , Oligonucleótidos , Fenómenos Magnéticos , Microbiología de AlimentosRESUMEN
Development of selective enrichment materials for the accurate analysis of ochratoxin a (OTA) in environmental and food samples is an effective way to protect human health. Here, a molecularly imprinted polymer (MIP) known as plastic antibody was synthesized onto the magnetic inverse opal photonic crystal microsphere (MIPCM) using a low-cost dummy template imprinting strategy targeting OTA. The MIP@MIPCM exhibited ultrahigh selectivity with an imprinting factor of 130, high specificity with cross-reactivity factors of 3.3-10.5, and large adsorption capacity of 60.5 µg/mg. Such MIP@MIPCM was used for selective capture of OTA in real samples which was quantified in combination with high-performance liquid chromatography, giving a wide linear detection range of 5-20,000 ng/mL, a detection limit of 0.675 ng/mL, and good recovery rates of 84-116%. Moreover, the MIP@MIPCM can be produced simply and rapidly and is very stable under different environmental conditions and easy to store and transport, so it is an ideal substitute of biological antibody modified materials for the selective enrichment of OTA in real samples.
Asunto(s)
Impresión Molecular , Polímeros Impresos Molecularmente , Humanos , Impresión Molecular/métodos , Microesferas , Polímeros/química , Cromatografía Líquida de Alta Presión , Adsorción , Fenómenos MagnéticosRESUMEN
Rapid identification and quantitative simultaneous analysis for multiple pesticide in real samples based on surface-enhanced Raman spectroscopy (SERS) is still a challenge because of sample complexity, reproducibility, and stability of SERS substrate. With use of colloidal silver nanoparticles loaded three-dimensional (3D) silica photonic microspheres (SPMs) array as the analytical platform, a SERS-based array assay for multiple pesticides was developed in this work. The silver nanoparticles were fixed into the gaps formed by the self-assembled nanospheres of the 3D SPMs to produce "hot spots", on which the Raman enhanced effect was up to 9.86 × 107 and the maximum electric field enhancement effect reached to 9.75 times, ensuring the target pesticides on the surface of the SERS-substrate integrated SPM can be detected sensitively. Using 2,4-dichlorophenoxyacetic acid (2,4-D), glyphosate, and imidacloprid as the testing pesticides, the label-free and high-throughput SERS assay for simultaneous detection of the pesticides was established, giving good linear detection ranges (0.1-204.8 µg/mL for 2,4-D, 0.3-247.9 µg/mL for glyphosate, and 0.2-204.8 µg/mL for imidacloprid) and low detection limits (3.03 ng/mL for 2,4-D, 3.14 ng/mL for glyphosate, and 8.82 ng/mL for imidacloprid). The spiked recovery rates in the real samples were measured in the range of 82-112%, which was consistent with that of the classical standard methods. The label-free 3D SERS array analytical platform provides a powerful tool for high-throughput and low-cost screening of multiple pesticide residues in real samples.
Asunto(s)
Nanopartículas del Metal , Plaguicidas , Plaguicidas/análisis , Nanopartículas del Metal/química , Dióxido de Silicio , Microesferas , Reproducibilidad de los Resultados , Plata/química , Espectrometría Raman/métodos , Ácido 2,4-DiclorofenoxiacéticoRESUMEN
We have demonstrated the proof-of-concept of a label-free fluorescence quantitative detection platform based on gold nanoparticle (AuNP) enhancement intrinsic fluorescence of protein on the silica photonic crystal microsphere (SPCM) array. The label-free one-step competitive fluorescence immunoassay protocol has been proposed on the surface of the SPCM. Aflatoxin B1 (AFB1) as a model molecule was detected by the newly established method. AFB1-bovine serum albumin and monoclonal antibodies (Abs) of anti-AFB1 have been immobilized on the surfaces of SPCMs and AuNPs, respectively. AuNPs remarkably enhanced the intrinsic fluorescence of artificial antigens on the surface of the SPCM at near UV excitation. The simulation of electric field distribution showed that the maximum value of the near-field enhancement |E/E0| of the SPCM with AuNPs could reach 20. The label-free fluorescence enhancement effect comes from the synergistic effects of photonic crystal effect and AuNP plasmon effect. Such a label-free fluorescence detection method can provide a linear detection range from 0.1 to 10 ng/mL with a limit of detection of 0.025 ng/mL and good specificity for AFB1. The recovery rates in the spiked cereal samples were measured in the range of 84.07 ± 5.71%-101.02 ± 5.13%, which were consistent with that of the traditional enzyme linked immunosorbent assay method. The label-free detection platform displays great application potential in biology, medicine, agriculture, food industry, chemical industry, energy source, and environmental protection.
Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , Microesferas , Oro/química , Dióxido de Silicio/química , Nanopartículas del Metal/química , Ensayo de Inmunoadsorción Enzimática , Aflatoxina B1/análisis , Límite de DetecciónRESUMEN
Chemical contaminants in food generally include natural toxins (mycotoxins, animal toxins, and phytotoxins), pesticides, veterinary drugs, environmental pollutants, heavy metals, and illegal additives. Developing a low-cost, simple, and rapid detection technology for harmful substances in food is urgently needed. Analytical methods based on different advanced materials have been developed into rapid detection methods for food samples. In particular, photonic crystal (PC) materials have a unique surface periodic structure, structural color, a large surface area, easy integration with photoelectronic and magnetic devices which have great advantages in the development of rapid, low-cost, and highly sensitive analytical methods. This review focuses on the PC materials in the view of their fabrication processes, functionalized recognition components for the specific recognition of hazardous substances, and applications in the separation, enrichment, and detection of chemical hazards in real samples. Suspension array based on three-dimensional PC microspheres by droplet-based microfluidic assembly is a great promising and powerful platform for food safety detection fields. For the PCs selective analysis, biological antibodies, aptamers, and molecularly imprinted polymers (MIPs) could be modified for specific recognition of target substances, particularly MIPs because of their low-cost and easy mass production. Based on these functional PCs, various toxic and hazardous substances can be selectively enriched or recognized in real samples and further quantified in combination of liquid chromatography method or optical detection methods including fluorescence, chemiluminescence, and Raman spectroscopy.
Asunto(s)
Impresión Molecular , Micotoxinas , Animales , Impresión Molecular/métodos , Polímeros/química , Inocuidad de los Alimentos , Sustancias PeligrosasRESUMEN
Development of an efficient detection method to monitor residual mycotoxins in food is very important to ensure food safety, but the complex food matrix seriously affects the detection sensitivity and accuracy. Here, using a three-dimensional ordered macroporous magnetic inverse photonic crystal microsphere (MPCM) as the supporting material, a molecularly imprinted polymer (MIP) that can selectively recognize aflatoxin B1 (AFB1) was synthesized through the dummy template imprinting strategy. The MPCM@MIP prepared by employing 5,7-dimethoxycoumarin as the template and methacrylic acid as the functional monomer displayed selectivity toward AFB1 (imprinting factor of 1.5) and could be used as a solid-phase extraction material. By coupling with high-performance liquid chromatography, an analytical method targeting AFB1 was established and displayed a wide linear range of 5-1000 ng/mL with a low detection limit of 0.4 ng/mL. The method showed a good recovery rate of 73-92% in AFB1-spiked soy sauce and vinegar samples. Moreover, the MPCM@MIP could be separated from the sample solution easily because of its magnetic performance, displaying a promising future not only in the enrichment of AFB1 to improve the detection sensitivity and accuracy but also in the removal of AFB1 from food and environmental samples.
Asunto(s)
Impresión Molecular , Polímeros Impresos Molecularmente , Adsorción , Aflatoxina B1/análisis , Cromatografía Líquida de Alta Presión , Fenómenos Magnéticos , Microesferas , Impresión Molecular/métodos , Polímeros/química , Extracción en Fase Sólida/métodosRESUMEN
A novel multiplex mycotoxin surface-enhanced Raman spectroscopy (SERS) immunoassay was established for the first time on different artificial antigen-modified silica photonic crystal microspheres (SPCMs), which can be integrated into a biochip array to achieve multiplex detection using corresponding antibody-functionalized gold nanoparticles (AuNPs) as the SERS nanotag. The unique optical structure of SPCMs is helpful to find the detection spots easily, accommodate a large amount of probe molecules, and enhance the Raman signal intensity. Such enhancement was confirmed by the simulation result, showing the electric field enhancing effect in SPCMs with AuNPs being 7 times. A competitive SERS immunoassay was established using antigen-modified SPCMs and mycotoxins to compete for binding antibody-functionalized SERS nanotags, displaying broad linear detection ranges of 0.001-0.1 ng/mL for aflatoxin B1 (AFB1), 0.01-10 ng/mL for ochratoxin A (OTA), and 0.001-0.1 ng/mL for zearalenone (ZEN) and low detection limits of 0.82 pg/mL for AFB1, 1.43 pg/mL for OTA, and 1.00 pg/mL for ZEN. In the spiked cereal samples, recovery rates of the method were measured in the range of 70.35-118.04% for the three mycotoxins, which was in agreement with that of the traditional enzyme-linked immunosorbent assay method. The SERS immunoassay for mycotoxin detection also showed high specificity and good repeatability and reproducibility. The new microsphere-based SERS immunoassay biochip only requires a one-step reaction and overcomes the disadvantages of fluorescence and chemiluminescence background signals. The work paves the way for further developing SERS-based microsphere suspension arrays for new targets.