Development of a rapid multiplexed assay for the direct screening of antimicrobial residues in raw milk.

Antimicrobial residues found to be present in milk can have both health and economic impacts. For these reasons, the widespread routine testing of milk is required. Due to delays with sample handling and test scheduling, laboratory-based tests are not always suited for making decisions about raw material intake and product release, especially when samples require shipping to a central testing facility. Therefore, rapid on-site screening tests that can produce results within a matter of minutes are required to facilitate rapid intake and product release processes. Such tests must be simple for use by non-technical staff. There is increasing momentum towards the development and implementation of multiplexing tests that can detect a range of important antimicrobial residues simultaneously. A simple in situ multiplexed planar waveguide device that can simultaneously detect chloramphenicol, streptomycin and desfuroylceftiofur in raw dairy milk, without sample preparation, has been developed. Samples are simply mixed with antibody prior to an aliquot being passed through the detection cartridge for 5 min before reading on a field-deployable portable instrument. Multiplexed calibration curves were produced in both buffer and raw milk. Buffer curves, for chloramphenicol, streptomycin and desfuroylceftiofur, showed linear ranges (inhibitory concentration (IC)20-IC80) of 0.1-0.9, 3-129 and 12-26 ng/ml, whilst linear range in milk was 0.13-0.74, 11-376 and 2-12 ng/ml, respectively, thus meeting European legislated concentration requirements for both chloramphenicol and streptomycin, in milk, without the need for any sample preparation. Desfuroylceftiofur-contaminated samples require only simple sample dilution to bring positive samples within the range of quantification. Assay repeatability and reproducibility were lower than 12 coefficient of variation (%CV), whilst blank raw milk samples (n = 9) showed repeatability ranging between 4.2 and 8.1%CV when measured on all three calibration curves. Graphical Abstract MBio SnapEsi reader and cartridge.

An evaluation of the capability of a biolayer interferometry biosensor to detect low-molecular-weight food contaminants.

The safety of our food is an essential requirement of society. One well-recognised threat is that of chemical contamination of our food, where low-molecular-weight compounds such as biotoxins, drug residues and pesticides are present. Low-cost, rapid screening procedures are sought to discriminate the suspect samples from the population, thus selecting only these to be forwarded for confirmatory analysis. Many biosensor assays have been developed as screening tools in food contaminant analysis, but these tend to be electrochemical, fluorescence or surface plasmon resonance based. An alternative approach is the use of biolayer interferometry, which has become established in drug discovery and life science studies but is only now emerging as a potential tool in the analysis of food contaminants. A biolayer interferometry biosensor was assessed using domoic acid as a model compound. Instrument repeatability was tested by simultaneously producing six calibration curves showing replicate repeatability (n = 2) ranging from 0.1 to 6.5 % CV with individual concentration measurements (n = 12) ranging from 4.3 to 9.3 % CV, giving a calibration curve midpoint of 7.5 ng/ml (2.3 % CV (n = 6)). Reproducibility was assessed by producing three calibration curves on different days, giving a midpoint of 7.5 ng/ml (3.4 %CV (n = 3)). It was further shown, using assay development techniques, that the calibration curve midpoint could be adjusted from 10.4 to 1.9 ng/ml by varying assay parameters before the simultaneous construction of three calibration curves in matrix and buffer. Sensitivity of the assay compared favourably with previously published biosensor data for domoic acid.

The potential of handheld near infrared spectroscopy to detect food adulteration: Results of a global, multi-instrument inter-laboratory study.

Fraud in the food supply system will be exacerbated by shortages caused by climate change and COVID-19's impact. The dried herbs market exemplifies complex supply chains attractive to criminals seeking financial gain. Real-time remote testing is achievable through development of globally accessible chemometric models for portable near infrared devices, deployed throughout supply chains. This study describes building of models for detection of oregano adulteration, on portable near infrared devices, and comparison to a laboratory-based Fourier-Transform Infrared spectroscopy method. 33/34 portable devices were able to correctly classify 5 out of 6 samples successfully with all adulterated samples being correctly classified following the use of appropriate transferability pre-processing routines. The devices native setup shows limited ability to perform a true screening of oregano using the setup offered. However modifications to the setup could in the future offer a solution that facilitates fit-for-purpose real time detection of adulterated samples within the supply chain.

A rapid food chain approach for authenticity screening: The development, validation and transferability of a chemometric model using two handheld near infrared spectroscopy (NIRS) devices.

This study assesses the application of a handheld, near infrared spectroscopy (NIRS) device, namely the NeoSpectra Micro, for the determination of oregano authenticity. Utilising a large sample set of oregano (n = 295) and potential adulterants of oregano (n = 109), models were developed and validated using SIMCA 15 software. The models demonstrated excellent predictability for the determination of authentic oregano and adulterant samples. The optimal model resulted in a 93.0% and 97.5% correct prediction for oregano and adulterants, respectively. Different standardisation approaches were assessed to determine model transferability to a second NIRS device. In the case of the second device, the best predictions were achieved with data that had not undergone any spectral standardisation (raw). Subsequently, the optimal model was able to correctly predict 90% of authentic oregano samples and 100% of the adulterant samples on the second device. This study demonstrates the potential of the device to be used as a simple, cost effective, reliable and handheld screening tool for the determination of oregano authenticity, at various stages of the food supply chain. It is believed that such forms of monitoring could be highly beneficial in other areas of food authenticity analysis to help combat the negative economical and health implications of food fraud.

Development of a rapid low cost fluorescent biosensor for the detection of food contaminants.

A prototype fluorescent based biosensor has been developed for the antibody based detection of food related contaminants. Its performance was characterised and showed a typical antibody binding signal of 200-2000 mV, a short term noise of 9.1 mV, and baseline slope of -0.016 mV/s over 4h. Bulk signal detection repeatability (n=23) and reproducibility (n=3) were less than 2.4%CV. The biosensor detection unit was evaluated using two food related model systems proving its ability to monitor both binding using commercial products and inhibition through the development of an assay. This assay development potential was evaluated by observing the biosensor's performance whilst appraising several labelled antibody and glass slide configurations. The molecular interaction between biotin and an anti-biotin antibody was shown to be inhibited by 41% due to the presence of biotin in a sample. A food toxin (domoic acid) calibration curve was produced, with %CVs ranging from 2.7 to 7.8%, and a midpoint of approximately 17 ng/ml with further optimisation possible. The ultimate aim of this study was to demonstrate the working principles of this innovative biosensor as a potential portable tool with the opportunity of interchangeable assays. The biosensor design is applicable for the requirements of routine food contaminant analysis, with respect to performance, functionality and cost.