Development of a novel duplex lateral flow test for simultaneous detection of casein and β-lactoglobulin in food

Galán-Malo, Patricia; Pellicer, Silvia; Pérez, María D.; Sánchez, Lourdes; Razquín, Pedro; Mata, Luis

doi:10.1016/j.foodchem.2019.04.039

Cited by 45 publications

(16 citation statements)

References 33 publications

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“…By increasing the sample volume to 25 µL (thus concentrating the sample 25 × compared with the 1 µL sample volume) LODs of 0.1 and 0.5 ppm for hazelnut and peanut were reached respectively (see Figure 3B). Despite the assay sensitivity improving with the increased sample volume, with these conditions at concentrations of 100 ppm and higher, a reduction of the intensity of the upper line (hazelnut) could be observed, as has been witnessed by Galan-Malo et al [24]. Although this was not considered a false negative, as three distinct lines were still clearly visible, it did warrant further exploration into the extent of the hook-effect in more concentrated samples.…”

Section: Multiplex Lateral Flow Immunoassaymentioning

confidence: 78%

“…However, this past year has seen an increase in the development of multiplex food allergen detection LFIAs, with the development of an assay for the detection of hazelnut, ovalbumin and casein in bakery products within 10 min [23]. A further example is the multiplex, low-ppm detection of both β-lactoglobulin and β-casein, two major allergenic milk proteins, within 10 min [24].…”

Section: Introductionmentioning

confidence: 99%

“…The hook-effect is a phenomenon that is commonly encountered in one-step, sandwich format LFIAs. It occurs where the free analyte and the analyte which is bound to a labeled antibody compete for the limited number of binding sites available on immobilized capture antibodies, leading to a reduction in colorimetric signal and sometimes false negative results [24,35,36]. Therefore, if the correct assay working range is not determined, it could lead to consumers erroneously believing a food with a high allergen content is safe.…”

Section: Introductionmentioning

confidence: 99%

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A Critical Comparison between Flow-through and Lateral Flow Immunoassay Formats for Visual and Smartphone-Based Multiplex Allergen Detection

2019

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(1) Background: The lack of globally standardized allergen labeling legislation necessitates consumer-focused multiplexed testing devices. These should be easy to operate, fast, sensitive and robust. (2) Methods: Herein, we describe the development of three different formats for multiplexed food allergen detection, namely active and passive flow-through assays, and lateral flow immunoassays with different test line configurations. (3) Results: The fastest assay time was 1 min, whereas even the slowest assay was within 10 min. With the passive flow approach, the limits of detection (LOD) of 0.1 and 0.5 ppm for total hazelnut protein (THP) and total peanut protein (TPP) in spiked buffer were reached, or 1 and 5 ppm of THP and TPP spiked into matrix. In comparison, the active flow approach reached LODs of 0.05 ppm for both analytes in buffer and 0.5 and 1 ppm of THP and TPP spiked into matrix. The optimized LFIA configuration reached LODs of 0.1 and 0.5 ppm of THP and TPP spiked into buffer or 0.5 ppm for both analytes spiked into matrix. The optimized LFIA was validated by testing in 20 different blank and spiked matrices. Using device-independent color space for smartphone analysis, two different smartphone models were used for the analysis of optimized assays.

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Section: Multiplex Lateral Flow Immunoassaymentioning

confidence: 78%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Critical Comparison between Flow-through and Lateral Flow Immunoassay Formats for Visual and Smartphone-Based Multiplex Allergen Detection

2019

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“…Still, it is vital to understand that highly concentrated samples can yield paradoxically low signal intensities, which could easily be misinterpreted by a consumer. However, it is reported that sample dilution (DF x 10–100) can minimize the occurrence of false-negatives [ [11] , [12] , [13] ]. While sample dilution is a prerequisite for allergen analysis, we cannot expect the consumer to do this.…”

Section: Resultsmentioning

confidence: 99%

“…Even integrated systems often require pre-treatment [ 10 ] or heat-assisted actuation to extract proteins into a testable liquid [ 3 ]. Finally, excluding a few multiplex LFIAs [ [11] , [12] , [13] ], allergen LFIAs are restricted to singleplex detection, which is limiting for individuals with co-existing allergies. Sample preparation is a major issue; indubitably, consumers do not have the laboratory skills required for extracting, pipetting, and diluting samples, and fully integrated analytical systems have so far mainly been developed for DNA-based analysis [ [14] , [15] , [16] ].…”

Section: Introductionmentioning

confidence: 99%

Interconnectable solid-liquid protein extraction unit and chip-based dilution for multiplexed consumer immunodiagnostics

Ross

Filippini

Nielen

et al. 2020

Analytica Chimica Acta

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While consumer-focused food analysis is upcoming, the need for multiple sample preparation and handling steps is limiting. On-site and consumer-friendly analysis paradoxically still requires laboratory-based and skill-intensive sample preparation methods. Here, we present a compact, inexpensive, and novel prototype immunosensor combining sample preparation and on-chip reagent storage for multiplex allergen lateral flow immunosensing. Our comprehensive approach paves the way for personalized consumer diagnostics. The prototype allows for handheld solid-liquid extraction, pipette-free on-chip dilution, and adjustment of sample concentrations into the appropriate assay dynamic working range. The disposable and interconnectable homogenizer unit allows for the extraction and 3D-sieve based filtration of allergenic proteins from solid bakery products in 1 min. The homogenizer interconnects with a 3D-printed unibody lab-on-a-chip (ULOC) microdevice, which is used to deliver precise volumes of sample extract to a reagent reservoir. The reagent reservoir is implemented for on-chip storage of carbon nanoparticle labeled antibodies and running buffer for dilution. The handheld prototype allows for total homogenization of solid samples, solid-liquid protein extraction, 3D-printed sieve based filtration, ULOC-enabled dilution, mixing, transport, and smartphone-based detection of hazelnut and peanut allergens in solid bakery products with limited operational complexity. The multiplex lateral flow immunoassay (LFIA) detects allergens as low as 0.1 ppm in real bakery products, and the system is already consumer-operable, demonstrating its potential for future citizen science approaches. The designed system is suitable for a wide range of analytical applications outside of food safety, provided an LFIA is available.

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Electrochemical Detection of β‐Lactoglobulin Allergen Using Titanium Dioxide/Carbon Nanochips/Gold Nanocomposite‐based Biosensor

et al. 2021

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A label-free electrochemical immunosensor was developed for the ultra-sensitive detection of β-lactoglobulin (β-LG). The novel nanocomplex of carbon nanochips, colloidal gold nanoparticles and titanium dioxide nanoparticles TiO 2 /CNC/AuC were constructed on conducting polymer, chitosan, and were characterised using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX). This nanocomplex interface was studied using cyclic voltammetry (CV) and showed great improvement at the gold electrode surface with enhanced electrochemical performance, sensitivity and selectivity for β-lactoglobulin. Under optimal parameters, the square wave voltammetry (SWV) response curve was determined from 0.01 pg/mL to 500 pg/mL using [Fe(CN) 6 ] 3À /4À ] redox probe. The calibration plot illustrates a linear relationship between log β-LG concentration and SWV current, with the limit of detection determined to be 0.01 pg/mL. This immunosensor displayed high sensitivity, selectivity, reproducibility and stability, and can be utilised for the detection of β-LG in real food samples.

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Development of a novel duplex lateral flow test for simultaneous detection of casein and β-lactoglobulin in food

Cited by 45 publications

References 33 publications

A Critical Comparison between Flow-through and Lateral Flow Immunoassay Formats for Visual and Smartphone-Based Multiplex Allergen Detection

A Critical Comparison between Flow-through and Lateral Flow Immunoassay Formats for Visual and Smartphone-Based Multiplex Allergen Detection

Interconnectable solid-liquid protein extraction unit and chip-based dilution for multiplexed consumer immunodiagnostics

Electrochemical Detection of β‐Lactoglobulin Allergen Using Titanium Dioxide/Carbon Nanochips/Gold Nanocomposite‐based Biosensor

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