Genomic identification and evolutionary analysis of chemosensory receptor gene families in two Phthorimaea pest species: insights into chemical ecology and host adaptation.

BACKGROUND: Insects rely on sophisticated sensitive chemosensory systems to sense their complex chemical environment. This sensory process involves a combination of odorant receptors (ORs), gustatory receptors (GRs) and ionotropic receptors (IRs) in the chemosensory system. This study focused on the identification and characterization of these three types of chemosensory receptor genes in two closely related Phthorimaea pest species, Phthorimaea operculella (potato tuber moth) and Phthorimaea absoluta (tomato leaf miner). RESULTS: Based on manual annotation of the genome, we identified a total of 349 chemoreceptor genes from the genome of P. operculella, including 93 OR, 206 GR and 50 IR genes, while for P. absoluta, we identified 72 OR, 122 GR and 46 IR genes. Through phylogenetic analysis, we observed minimal differences in the number and types of ORs and IRs between the potato tuber moth and tomato leaf miner. In addition, we found that compared with those of tomato leaf miners, the gustatory receptor branch of P. operculella has undergone a large expansion, which may be related to P. absoluta having a narrower host range than P. operculella. Through analysis of differentially expressed genes (DEGs) of male and female antennae, we uncovered 45 DEGs (including 32ORs, 9 GRs, and 4 IRs). CONCLUSIONS: Our research provides a foundation for exploring the chemical ecology of these two pests and offers new insights into the dietary differentiation of lepidopteran insects, while simultaneously providing molecular targets for developing environmentally friendly pest control methods based on insect chemoreception.

Elucidating the Bioinspired Synthesis Process of Magnetosomes-Like Fe3O4 Nanoparticles.

Iron oxide nanoparticles are a kind of important biomedical nanomaterials. Although their industrial-scale production can be realized by the conventional coprecipitation method, the controllability of their size and morphology remains a huge challenge. In this study, a kind of synthetic polypeptide Mms6-28 which mimics the magnetosome protein Mms6 is used for the bioinspired synthesis of Fe3O4 nanoparticles (NPs). Magnetosomes-like Fe3O4 NPs with uniform size, cubooctahedral shape, and smooth crystal surfaces are synthesized via a partial oxidation process. The Mms6-28 polypeptides play an important role by binding with iron ions and forming nucleation templates and are also preferably attached to the [100] and [111] crystal planes to induce the formation of uniform cubooctahedral Fe3O4 NPs. The continuous release and oxidation of Fe2+ from pre-formed Fe2+-rich precursors within the Mms6-28-based template make the reaction much controllable. The study affords new insights into the bioinspired- and bio-synthesis mechanism of magnetosomes.

Synthetic biology for the food industry: advances and challenges.

As global environmental pollution increases, climate change worsens, and population growth continues, the challenges of securing a safe, nutritious, and sustainable food supply have become enormous. This has led to new requirements for future food supply methods and functions. The use of synthetic biology technology to create cell factories suitable for food industry production and renewable raw material conversion into: important food components, functional food additives, and nutritional chemicals, represents an important method of solving the problems faced by the food industry. Here, we review the recent progress and applications of synthetic biology in the food industry, including alternatives to: traditional (artificial pigments, meat, starch, and milk), functional (sweeteners, sugar substitutes, nutrients, flavoring agents), and green (green fiber, degradable packing materials, green packaging materials and food traceability) foods. Furthermore, we discuss the future prospects of synthetic biology-based applications in the food industry. Thus, this review may serve as a reference for research on synthetic biology in the: food safety, food nutrition, public health, and health-related fields.

Rapid detection of melamine by DNA Walker mediated SERS sensing technique based on signal amplification function.

A new method is proposed for detecting typical melamine dopants in food using surface-enhanced Raman scattering (SERS) biosensing technology. Melamine specific aptamer was used as the identification probe, and gold magnets (AuNPs@MNPs) and small gold nanoparticles (AuNPs@MBA) were used as the basis for Raman detection. The Raman signal of the detection system can directly detect melamine quantitatively. Under optimized conditions, the detection of melamine was carried out in the low concentration range of 0.001-500 mg/kg, the enhancement factor (EF) was 2.3 × 107, and the detection limit was 0.001 mg/kg. The method is sensitive and rapid, and can be used for the rapid detection of melamine in the field environment.

Application of DNA-fueled molecular machines in food safety testing.

Food is consumed by humans, which is indispensable to human life. Therefore, considerable attention of the whole society has been paid to food safety. Over the last few years, dramatic social development has brought new challenges to food safety, making developing new and quick methods for on-site food safety testing an important necessity. As a result, DNA-fueled molecular machines, characterized by high efficiency, accuracy, and sensitivity in testing, have come into the spotlight, based on which sensors can be constructed to detect toxic and harmful substances in food products. This study reviewed recent research on several DNA-fueled molecular machines, including DNA tweezers, DNA walkers, and DNA origami, for rapidly detecting toxic and harmful substances. Based on the above studies, the sensitivity and timeliness of several DNA molecular machines were summarized and compared, and the development prospect of DNA fuel molecular machines in the field of food safety detection was prospected.

Development and perspectives of rapid detection technology in food and environment.

Food safety become a hot issue currently with globalization of food trade and food supply chains. Chemical pollution, microbial contamination and adulteration in food have attracted more attention worldwide. Contamination with antibiotics, estrogens and heavy metals in water environment and soil environment have also turn into an enormous threat to food safety. Traditional small-scale, long-term detection technologies have been unable to meet the current needs. In the monitoring process, rapid, convenient, accurate analysis and detection technologies have become the future development trend. We critically synthesizing the current knowledge of various rapid detection technology, and briefly touched upon the problem which still exist in research process. The review showed that the application of novel materials promotes the development of rapid detection technology, high-throughput and portability would be popular study directions in the future. Of course, the ultimate aim of the research is how to industrialization these technologies and apply to the market.

CRISPR/Cas12a-based technology: A powerful tool for biosensing in food safety.

BACKGROUND: In the context of the current pandemic caused by the novel coronavirus, molecular detection is not limited to the clinical laboratory, but also faces the challenge of the complex and variable real-time detection fields. A series of novel coronavirus events were detected in the process of food cold chain packaging and transportation, making the application of molecular diagnosis in food processing, packaging, transportation, and other links urgent. There is an urgent need for a rapid detection technology that can adapt to the diversity and complexity of food safety. SCOPE AND APPROACH: This review introduces a new molecular diagnostic technology-biosensor analysis technology based on CRISPR-Cas12a. Systematic clarification of its development process and detection principles. It summarizes and systematically organizes its applications in viruses, food-borne pathogenic bacteria, small molecule detection, etc. In the past four years, which provides a brand-new and comprehensive solution for food detection. Finally, this article puts forward the challenges and the prospects for food safety. KEY FINDINGS AND CONCLUSIONS: The novel coronavirus hazards infiltrated every step of the food industry, from processing to packaging to transportation. The biosensor analytical technology based on CRISPR-Cas12a has great potential in the qualitative and quantitative analysis of infectious pathogens. CRISPR-Cas12a can effectively identify the presence of the specific nucleic acid targets and the small changes in sequences, which is particularly important for nucleic acid identification and pathogen detection. In addition, the CRISPR-Cas12a method can be adjusted and reconfigured within days to detect other viruses, providing equipment for nucleic acid diagnostics in the field of food safety. The future work will focus on the development of portable microfluidic devices for multiple detection. Shao et al. employed physical separation methods to separate Cas proteins in different microfluidic channels to achieve multiple detection, and each channel simultaneously detected different targets by adding crRNA with different spacer sequences. Although CRISPR-Cas12a technology has outstanding advantages in detection, there are several technical barriers in the transformation from emerging technologies to practical applications. The newly developed CRISPR-Cas12a-based applications and methods promote the development of numerous diagnostic and detection solutions, and have great potential in medical diagnosis, environmental monitoring, and especially food detection.

Superhydrophobic Paper-Based Microfluidic Field-Effect Transistor Biosensor Functionalized with Semiconducting Single-Walled Carbon Nanotube and DNAzyme for Hypocalcemia Diagnosis.

Hypocalcemia is caused by a sharp decline in blood calcium concentration after dairy cow calving, which can lead to various diseases or even death. It is necessary to develop an inexpensive, easy-to-operate, reliable sensor to diagnose hypocalcemia. The cellulose-paper-based microfluidic field-effect biosensor is promising for point-of-care, but it has poor mechanical strength and a short service life after exposure to an aqueous solution. Octadecyltrichlorosilane (OTS), as a popular organosilane derivative, can improve the hydrophobicity of cellulose paper to overcome the shortage of cellulose paper. In this work, OTS was used to produce the superhydrophobic cellulose paper that enhances the mechanical strength and short service life of MFB, and a microfluidic field-effect biosensor (MFB) with semiconducting single-walled carbon nanotubes (SWNTs) and DNAzyme was then developed for the Ca2+ determination. Pyrene carboxylic acid (PCA) attached to SWNTs through a non-covalent π-π stacking interaction provided a carboxyl group that can bond with an amino group of DNAzyme. Two DNAzymes with different sensitivities were designed by changing the sequence length and cleavage site, which were functionalized with SPFET/SWNTs-PCA to form Dual-MFB, decreasing the interference of impurities in cow blood. After optimizing the detecting parameters, Dual-MFB could determine the Ca2+ concentration in the range of 25 µM to 5 mM, with a detection limit of 10.7 µM. The proposed Dual-MFB was applied to measure Ca2+ concentration in cow blood, which provided a new method to diagnose hypocalcemia after dairy cow calving.

The orphan nuclear receptor Nur77 plays a vital role in BPA-induced PC12 cell apoptosis.

Bisphenol A (BPA) is a typical environmental endocrine disruptor that can migrate into organisms through skin contact, breathing, diet and various other approaches. The reproductive toxicity and neurotoxicity of BPA has been confirmed by several toxicological studies. However, the neurotoxicity of BPA is still controversial. In the present study, we used PC12 cells as a model to investigate the mechanism of BPA-induced neuronal apoptosis. BPA exposure reduced cell viability, altered cell morphology and aggravated intracellular Lactate dehydrogenase (LDH) release, intracellular Ca2+ concentration, Reactive oxygen species (ROS) levels, apoptosis and the reduction in the mitochondrial transmembrane potential (ΔΨm). Moreover, the results of the Western blot (WB) and Real-time quantitative polymerase chain reaction (RT-qPCR) assays indicated that the expression levels of Nur77 in the BPA group were down-regulated and accompanied by the downregulation of the NF-κb/Bcl-2 proteins and the upregulation of cleaved-caspase 3, which is a marker of apoptosis. However, these changes were significantly reversed with the upregulation of the Nur77 protein by introducing plasmids carrying the nur77 gene. These results indicated that BPA-induced apoptosis was closely related to Nur77-mediated inhibition of the NF-κb/Bcl-2 pathway.

Surface-enhanced Raman spectroscopy aptasensor for simultaneous determination of ochratoxin A and zearalenone using Au@Ag core-shell nanoparticles and gold nanorods.

The design and fabrication of a surface-enhanced Raman scattering (SERS) aptasensor for simultaneous detection of zearalenone (ZEN) and ochratoxin A (OTA) in wheat and corn samples is described. The capture and reporter probes were SH-cDNA-modified gold nanorods and SH-Apt-modified Au@Ag core-shell nanoparticles, respectively. After recognizing OTA and ZEN aptamers and complementary strands (SH-cDNA), the reporter probe generated a strong SERS signal. The preferred binding of OTA and ZEN aptamers to OTA and ZEN, respectively, caused reporter probes to release the capture probes, resulting in a linear decrease in SERS intensity. The detection of OTA showed good linearity with an R2 value of 0.986, which could be maintained across a wide concentration range (0.01 to 100 ng/mL), with the limit of detection of 0.018 ng/mL. For detection of ZEN, good linearity with an R2 value of 0.987 could be maintained across a wide concentration range (0.05 to 500 ng/mL), with 0.054 ng/mL as the limit of detection. Good accuracy (relative standard deviation < 4.2%) during mycotoxin determination as well as excellent quantitative recoveries (96.0-110.7%) during the analysis of spiked real samples was achieved. The proposed SERS aptasensor exhibited excellent performance in the detection of OTA and ZEN in real food samples. Hence, by simply changing the aptamer, this new model can be applied to the detection of multiple mycotoxins in the food industry.

Vertical Flow Assay for Inflammatory Biomarkers Based on Nanofluidic Channel Array and SERS Nanotags.

There is a great demand for the development of detection assays for inflammation infection diagnosis with high throughput and ultrasensitivity. Herein, a vertical flow assay system with functionalized nanoporous anodic aluminum oxide (AAO) as sensing membrane, and encoded core-shell surface enhanced Raman scattering (SERS) nanotags as labels for multiple inflammatory biomarkers detection is presented. A 2 × 2 test array on the porous AAO is developed and modified with multiple capture antibodies to capture inflammatory biomarkers from samples. Due to the high surface area to volume ratio of the AAO membrane, and its influence on plasmonic coupling, the electromagnetic field of the encoded core-shell SERS nanotags is enhanced. Detection limits of 53.4, 4.72, 48.3, and 7.53 fg mL-1 are realized for C reactive protein, interleukin-6, serum amyloid A, and procalcitonin, respectively, with a linear dynamic range spanning at least five orders of magnitude. In addition, the proposed method also shows acceptable accuracy and repeatability for the detection of clinical samples. Therefore, this approach is expected to be a powerful point of care testing tool for disease diagnosis in facility limited areas.

Plasmon-coupled microcavity aptasensors for visual and ultra-sensitive simultaneous detection of Staphylococcus aureus and Escherichia coli.

Septicemia and bacteremia are serious infections in the bloodstream. Thus, time-saving and ultra-sensitive pathogenic bacteria detection is highly required. Herein, we constructed gold nanoparticle-modified polystyrene microspheres (Au/PS) as plasmon-coupled microcavities to realize simultaneous detection of Staphylococcus aureus and Escherichia coli based on a fluorescence and surface-enhanced Raman spectroscopy (SERS) dual-mode method. Fluorescence imaging, serving as a means for assistant validation and rapid screening, was carried out to achieve qualitative and semi-quantitative determination, which gave us visual information of the existence and distribution of the target bacteria. Meanwhile, SERS test was conducted to realize ultra-sensitive quantitative detection. The evanescent wave aroused from total internal reflection in PS microcavities coupled with the localized electromagnetic field from surface plasmons of gold nanoparticles to improve light-matter interaction synergistically, leading to an enhancement factor of 2.25 × 1011 for SERS sensing. The whole measurement was carried out in a typical sandwich assay of "capture probe-target bacteria-signal probe." As a result, calibrated concentration response curves demonstrated the sensitive quantitative detection with the limit of detection (LOD) of 3 cfu/mL for S. aureus and 2 cfu/mL for E. coli. This rapid, ultra-sensitive, and visual sensing method was further developed for dual-bacteria detection in the whole blood samples.

Vertical flow assays based on core-shell SERS nanotags for multiplex prostate cancer biomarker detection.

Rapid, simultaneous, and sensitive quantification of multiplex prostate biomarkers plays an important role in early diagnosis, especially for obese men and patients. Herein, a surface-enhanced Raman scattering (SERS)-based vertical flow assay (VFA) is presented for simultaneous detection of multiplex prostate cancer biomarkers, such as prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), and alpha-fetoprotein (AFP) on a single test spot. In practice, Raman dyes (RDs) encoded core-shell SERS nanotags instead of conventional gold colloids used in the colorimetric assay are employed in the sensing membrane of SERS based VFAs for multiplex protein detection. Because of the enhanced Raman signal of the core-shell nanostructure and the high surface area to volume ratio (SVR) of the porous sensing membrane, this proposed biosensor shows a wide linear dynamic range (LDR) with detection limits of 0.37, 0.43, and 0.26 pg mL-1 for PSA, CEA, and AFP, respectively, suggesting that this approach can be a good candidate in point of care testing (POCT) for rapid and sensitive biomarker detection and has a huge potential in multiplex analysis and cancer diagnosis.

A Non-Destructive and Direction-Insensitive Method Using a Strain Sensor and Two Single Axis Angle Sensors for Evaluating Corn Stalk Lodging Resistance.

Corn stalk lodging is caused by different factors, including severe wind storms, stalk cannibalization, and stalk rots, and it leads to yield loss. Determining how to rapidly evaluate corn lodging resistance will assist scientists in the field of crop breeding to understand the contributing factors in managing the moisture, chemical fertilizer, and weather conditions for corn growing. This study proposes a non-destructive and direction-insensitive method, using a strain sensor and two single axis angle sensors to measure the corn stalk lodging resistance in the field. An equivalent force whose direction is perpendicular to the stalk is utilized to evaluate the corn lodging properties when a pull force is applied on the corn stalk. A novel measurement device is designed to obtain the equivalent force with the coefficient of variation (CV) of 4.85%. Five corn varieties with two different planting densities are arranged to conduct the experiment using the novel measurement device. The experimental results show that the maximum equivalent force could reach up to 44 N. A strong relationship with the square of the correlation coefficient of 0.88 was obtained between the maximum equivalent forces and the corn field’s stalk lodging rates. Moreover, the stalk lodging angles corresponding to the different pull forces over a measurement time of 20 s shift monotonically along the equivalent forces. Thus, the non-destructive and direction-insensitive method is an excellent tool for rapid analysis of stalk lodging resistance in corn, providing critical information on in-situ lodging dynamics.

Reflectance and fluorescence characterization of maize species using field laboratory measurements and lidar remote sensing.

Laser-induced fluorescence is an important technique to study photosynthesis and plants. Information on chlorophyll and other pigments can be obtained. We have been using a mobile laboratory in a Chinese experimental farm setting to study maize (Zea mays L.) leaves by reflectance and fluorescence measurements and correlated the spectroscopic signals to the amount of fertilizer supplied. Further, we studied five different species of maize using the remote monitoring of the fluorescence signatures obtained with the same mobile laboratory, but now in a laser radar remote-sensing configuration. The system separation from the target area was 50 m, and 355 nm pulsed excitation using the frequency-tripled output from an Nd:YAG laser was employed. Principal component analysis and linear discriminant analysis were combined to identify the different maize species using their fluorescence spectra. Likewise, the spectral signatures in reflectance and fluorescence frequently allowed us to separate different fertilizer levels applied to plants of the same species.

A Low Cost Compact Measurement System Constructed Using a Smart Electrochemical Sensor for the Real-Time Discrimination of Fruit Ripening.

Ethylene as an indicator for evaluating fruit ripening can be measured by very sensitive electrochemical gas sensors based on a high-resolution current produced by a bias potential applied to the electrodes. For this purpose, a measurement system for monitoring ethylene gas concentrations to evaluate fruit ripening by using the electrochemical ethylene sensor was successfully developed. Before the electrochemical ethylene sensor was used to measure the ethylene gas concentrations released from fruits, a calibration curve was established by the standard ethylene gases at concentrations of 2.99 ppm, 4.99 ppm, 8.01 ppm and 10 ppm, respectively, with a flow rate of 0.4 L·min(-1). From the calibration curve, the linear relationship between the responses and concentrations of ethylene gas was obtained in the range of 0-10 ppm with the correlation coefficient R² of 0.9976. The micropump and a novel signal conditioning circuit were implemented in this measurement, resulting in a rapid response in detecting ethylene concentrations down to 0.1 ppm in air and in under 50 s. In this experiment, three kinds of fruits-apples, pears and kiwifruits-were studied at a low concentration (under 0.8 ppm) of trace ethylene content in the air exhaled by fruits. The experimental results showed that a low cost, compact measurement system constructed by using an electrochemical ethylene sensor has a high sensitivity of 0.3907 V·ppm(-1) with a theoretical detection limit of 0.413 ppm, and is non-invasive and highly portable.

The Development of a Portable SPR Bioanalyzer for Sensitive Detection of Escherichia coli O157:H7.

The purpose of this study was to develop a portable surface plasmon resonance (SPR) bioanalyzer for the sensitive detection of Escherichia coli O157:H7 in comparison with an enzyme-linked immunosorbent assay (ELISA). The experimental setup mainly consisted of an integrated biosensor and a homemade microfluidic cell with a three-way solenoid valve. In order to detect Escherichia coli O157:H7 using the SPR immunoassay, 3-mercaptopropionic acid (3-MPA) was chemisorbed onto a gold surface via covalent bond for the immobilization of biological species. 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) were used as crosslinker reagents to enable the reaction between 3-MPA and Escherichia coli O157:H7 antibodies by covalent -CO-NH- amide bonding. The experimental results were obtained from the Escherichia coli O157:H7 positive samples prepared by 10-, 20-, 40-, 80-, and 160-fold dilution respectively, which show that a good linear relationship with the correlation coefficient R of 0.982 existed between the response units from the portable SPR bioanalyzer and the concentration of Escherichia coli O157:H7 positive samples. Moreover, the theoretical detection limit of 1.87 × 10³ cfu/mL was calculated from the positive control samples. Compared with the Escherichia coli O157:H7 ELISA kit, the sensitivity of this portable SPR bioanalyzer is four orders of magnitude higher than the ELISA kit. The results demonstrate that the portable SPR bioanalyzer could provide an alternative method for the quantitative and sensitive determination of Escherichia coli O157:H7 in field.

Considerations on Circuit Design and Data Acquisition of a Portable Surface Plasmon Resonance Biosensing System.

The aim of this study was to develop a circuit for an inexpensive portable biosensing system based on surface plasmon resonance spectroscopy. This portable biosensing system designed for field use is characterized by a special structure which consists of a microfluidic cell incorporating a right angle prism functionalized with a biomolecular identification membrane, a laser line generator and a data acquisition circuit board. The data structure, data memory capacity and a line charge-coupled device (CCD) array with a driving circuit for collecting the photoelectric signals are intensively focused on and the high performance analog-to-digital (A/D) converter is comprehensively evaluated. The interface circuit and the photoelectric signal amplifier circuit are first studied to obtain the weak signals from the line CCD array in this experiment. Quantitative measurements for validating the sensitivity of the biosensing system were implemented using ethanol solutions of various concentrations indicated by volume fractions of 5%, 8%, 15%, 20%, 25%, and 30%, respectively, without a biomembrane immobilized on the surface of the SPR sensor. The experiments demonstrated that it is possible to detect a change in the refractive index of an ethanol solution with a sensitivity of 4.99838 × 10(5) ΔRU/RI in terms of the changes in delta response unit with refractive index using this SPR biosensing system, whereby the theoretical limit of detection of 3.3537 × 10(-5) refractive index unit (RIU) and a high linearity at the correlation coefficient of 0.98065. The results obtained from a series of tests confirmed the practicality of this cost-effective portable SPR biosensing system.

Comparative transcriptome analysis of the antenna and proboscis reveals feeding state-dependent chemosensory genes in Eupeodes corollae.

The physiological state of an insect can affect its olfactory system. However, the molecular mechanism underlying the effect of nutrition-dependent states on odour-guided behaviours in hoverflies remains unclear. In this study, comparative transcriptome analysis of the antenna and proboscis from Eupeodes corollae under different feeding states was conducted. Compared with the previously published antennal transcriptome, a total of 32 novel chemosensory genes were identified, including 4 ionotropic receptors, 17 gustatory receptors, 9 odorant binding proteins and 2 chemosensory proteins. Analysis of differences in gene expression between different feeding states in male and female antennae and proboscises revealed that the expression levels of chemosensory genes were impacted by feeding state. For instance, the expression levels of EcorOBP19 in female antennae, EcorOBP6 in female proboscis, and EcorOR6, EcorOR14, EcorIR5 and EcorIR84a in male antennae were significantly upregulated after feeding. On the other hand, the expression levels of EcorCSP7 in male proboscis and EcorOR40 in male antennae were significantly downregulated. These findings suggest that nutritional state plays a role in the adaptation of hoverflies' olfactory system to food availability. Overall, our study provides important insights into the plasticity and adaptation of chemosensory systems in hoverflies.

Carboxyl-functionalized two-dimensional MXene-Au nanocomposites were prepared as SERS substrates for the detection of melamine in dairy products.

In the present study, we address the limitations of conventional surface-enhanced Raman scattering (SERS) techniques for sensitive and stable detection of melamine in food products, especially dairy. To overcome these challenges, we developed a novel SERS-active substrate by incorporating gold nanoparticles (AuNPs) onto carboxyl-functionalized two-dimensional (2D) MXene material doped with nitrides, specifically Au-Ti2N-COOH. Our strategy leverages the unique physicochemical properties of MXene, a class of atomically thin, 2D transition metal carbides/nitrides, with tunable surface functionalities. By modifying the MXene surface with AuNPs and introducing carboxyl groups (-COOH), we successfully enhanced the interaction between the substrate and melamine molecules. The carboxyl groups form hydrogen bonds with the amino groups on the melamine's triazine ring, facilitating the adsorption of melamine molecules within the 'hotspot' regions responsible for SERS signal amplification. A series of characterization methods were used to confirm the successful synthesis of Au-Ti2N-COOH composites.Using Au-Ti2N-COOH as the SERS substrate, we detected melamine in spiked dairy product samples with significantly enhanced sensitivity and stability compared to nitride-doped MXene alone. The detection limit in liquid milk stands at 3.7008 µg kg-1, with spike recovery rates ranging from 99.84% to 107.55% and an approximate RSD of 5%. This work demonstrates the effectiveness of our approach in designing a label-free, rapid, and robust SERS platform for the accurate quantitation of melamine contamination in food, thereby mitigating health risks associated with melamine adulteration.