Nanomaterial-Enhanced Biosensors for Detection of Salmonella typhimurium in Food Samples

Shivangi Kosma¹, Riya Ritika Singh², Manoj Kumar Patel^3*

^1,2,3 Nano-Biology Laboratory, School of Studies in Life Science, Pt. Ravishankar Shukla University, Raipur 492010, Chhattisgarh, India

Graphical Abstract:

Description: Diagram showing transmission of S. typhimurium from food samples and further detection of pathogen by biosensor.

Abstract

The rapid and accurate detection of Salmonella typhimurium remains a critical priority in food safety, environmental monitoring, and public health due to the limitations of conventional diagnostic methods. This review provides a comprehensive overview of recent advancements in nanomaterial-enabled biosensors, emphasizing their synthesis, application, and limitations in pathogen detection. Conventional techniques- including culture-based assays, serotyping, biochemical tests, immunological methods, and molecular diagnostic care discussed alongside their respective advantages and drawbacks. The integration of nanotechnology is highlighted as a transformative step, where nanomaterials such as graphene, metal oxides, carbon nanotubes, and quantum dots enhance biosensor sensitivity, selectivity, and portability through their unique physicochemical properties. Various biosensing platforms, including electrochemical, optical, piezoelectric, thermal, and DNA-based sensors, are examined with a focus on their mechanisms, analytical performance, and pathogen-specific innovations. Attention is given to nanomaterial-assisted DNA biosensors that achieve low limits of detection and rapid response times through efficient signal amplification and biorecognition strategies. The review also outlines essential drawbacks and limitations of biosensor development and how it affects reproducibility and performance reliability. An overview of multiplexed detection, real-time monitoring, CRISPR-integrated platforms, and point-of-care devices for next-generation diagnostic systems. Overall, the integration of nanotechnology with biosensor platforms offers a powerful pathway toward faster, more accurate and field-deployable detection of S. typhimurium and other infectious agents.

Keywords: Biosensor, DNA Biosensor, Food Pathogen, Nanomaterial, Salmonella typhimurium

1. Introduction

Biosensor devices detect analytes and convert their concentration into readable signals which are pivotal in medical diagnostics, physiological monitoring, and disease prevention (Li et al., 2023). These sophisticated analytical tools integrate biological recognition elements with signal transduction and amplification mechanisms, making them indispensable for monitoring biomarkers and enabling early disease detection (Patial et al., 2025). The inherent specificity of biomolecules, when coupled with the unique properties of nanomaterials, can facilitate the development of biosensors capable of real-time, non-invasive health monitoring (Mishra et al., 2025). Historically, the main cause of mortality has been infectious diseases. Infectious diseases have not surpassed other causes of death worldwide until recently. However, infectious diseases still affect people today, particularly in low-income nations. Additionally, the advent of novel diseases like COVID-19 and the reappearance of long-term illnesses pose a hazard to public health. The introduction of infections to new host populations, the evolution of antibiotic resistance, vaccine hesitancy, or the transfer of diseases from animals to people are frequently the causes of disease emergence. Infectious diseases also affect domesticated and wild plants and animals. Zoonotic pathogens, or infections that infect humans, are frequently found in animals and can lead to zoonotic diseases. Destructive infectious diseases affect both plants and animals, and some of these illnesses can have serious consequences (Rahman et al., 2020). The critical role of biosensors in detecting various diseases necessitates highly precise biomarkers, minimally invasive approaches, and meticulous differentiation among markers associated with diverse health conditions (Narware et al., 2025). The increasing demand for rapid and accurate detection methods across diverse fields, including healthcare, environmental analysis, and food safety, has driven significant interest in nanomaterial-enabled sensors (Pandey, 2022; Subhan et al., 2025). These biosensors offer superior sensitivity and specificity for detecting biological compounds, converting complex bioanalytical measurements into easily interpretable formats (Aftab et al., 2025). This enhanced capability is particularly crucial given the global burden of diseases such as ischemic heart disease, lung cancer, and cirrhosis, where early and accurate diagnosis is critical for effective treatment and improved patient outcomes (Pirzada & Altıntaş, 2019). The integration of nanotechnology with biosensors has paved the way for novel sensing mechanisms, significantly enhancing the performance and detection capabilities of existing biosensing platforms (Ramesh et al., 2022). This review delves into the synthesis, characterization, and diverse applications of nanomaterials in biosensing, highlighting recent advancements and challenges in the field. Specifically, it explores how the unique physicochemical properties of nanomaterials, such as their high surface-to-volume ratio and quantum effects, contribute to improved sensor sensitivity and selectivity. 

1.1. Transmission of Salmonella typhimurium

Salmonella typhimurium is primarily transmitted through the consumption of contaminated food or water; types are shown in Figure 1. Foods commonly associated with Salmonella contamination include raw or undercooked eggs, poultry, meat, dairy products, and fresh produce. Additionally, cross-contamination can occur through contact with infected animals or surfaces (Won & Lee, 2017).

Figure 1: Classification of Salmonella (Gram-Negative Bacteria)

1.2. Methods of Detection

Foodborne illnesses are extremely dangerous to human health and have a significant financial impact on the entire planet. Foodborne infections, including thrombotic thrombocytopenic purpura (TTP), hemorrhagic colitis, typhoid, acute gastroenteritis, diarrhea, and hemolytic uremic syndrome (HUS), are usually brought on by bacteria, viruses, and parasites that contaminate food from the point of harvesting until it is consumed. Therefore, to safeguard the food supply and prevent foodborne infections, it is imperative to identify foodborne pathogenic bacteria as soon as possible. Foodborne pathogen detection is associated with both traditional (i.e., culture-based, biochemical test-based, immunological-based, and nucleic acid-based procedures) and sophisticated (i.e., hybridization-based, array-based, spectroscopy-based, and biosensor-based process) techniques (Paranthaman et al., 2022). For industrial food applications, detection techniques could meet needs like non-labour intensiveness, efficiency, speed, specificity, sensitivity, and accuracy levels (Kabiraz et al., 2023).

1.3. Nanomaterial and Biosensing

The purpose of nanomaterial and biosensor development is multifaceted and encompasses various fields including healthcare, environmental monitoring, food safety, and security. In healthcare, Early Disease identification, Individualized Medicine, Point-of-Care Testing, Implanted Devices. In Environmental Observation: Pollution Detection, Water Quality Monitoring, Environmental Remediation. In Food Safety: Pathogen Detection, Quality Control, Traceability. In Security and Defense: Chemical and Biological Threat Detection, Biodefense (Bhalla et al., 2016). In summary, the development of nanomaterial-based biosensors serves diverse purposes ranging from healthcare diagnostics and environmental monitoring to food safety assurance and security applications, ultimately contributing to improved public health, safety, and quality of life.

2. Conventional Techniques

Conventional diagnostic techniques for the detection of Salmonella typhimurium have been well-established and widely used in laboratories (Cabral, 2010; Law et al., 2015).

2.1 Culture-Based Methods

These methods involve the cultivation of Salmonella on selective and differential media. Procedure involves (a) pre-Enrichment; Samples are incubated in non-selective broth to revive stressed cells. (b) Selective Enrichment, Samples are transferred to selective enrichment broths that favor the growth of Salmonella. (c) Selective Plating, Enriched samples are plated on selective agars like Xylose Lysine Deoxycholate (XLD) agar, Hekaton enteric agar, or Bismuth Sulphite agar. Identification and presumptive Salmonella colonies are further identified by biochemical tests and serotyping. Advantages- High specificity and ability to isolate and identify viable bacteria. Disadvantages- Time-consuming (can take 4-7 days), labor-intensive, and requires skilled personnel.

2.2. Serotyping

Identification of Salmonella serotypes based on antigen-antibody reactions. Procedure involves Slide Agglutination Test (Grimont & Weill, 2007). Uses antisera to detect specific O (somatic) and H (flagellar) antigens. Advantages- Provides specific serotype information, which is important for epidemiological studies. Disadvantages- Requires pure cultures and specific antisera.

2.3. Biochemical Tests

Utilizes the metabolic and enzymatic activities of Salmonella to identify the bacteria. Procedure involves Triple Sugar Iron (TSI) Agar- Differentiates based on carbohydrate fermentation and hydrogen sulphide production. (a) Urease Test- Detects the ability to hydrolyze urea. (b) Indole Test- Detects the production of indole from tryptophan (MacFaddin, 2000; Cheesbrough, 2006; Janda & Abbott, 2007). Advantages: Simple and inexpensive. Disadvantages- Requires pure cultures and multiple tests for accurate identification.

2.4. Molecular Methods

Polymerase Chain Reaction (PCR)- Amplifies specific DNA sequences unique to Salmonella. The procedure involves DNA extracting from samples. PCR is performed using specific primers targeting Salmonella genes (e.g., invA gene) (Rahn et al., 1992; Malorny et al., 2003). Advantages: High sensitivity, specificity, and rapid results (within hours). Disadvantages- Requires specialized equipment and expertise.

2.5. Immunological Methods

Enzyme-Linked Immunosorbent Assay (ELISA) involves detection of Salmonella antigens or antibodies using enzyme-labelled antibodies. Procedure involves Samples which are incubated with specific antibodies, Enzyme-conjugated secondary antibodies are added, Substrate is added to produce a measurable color change (Law et al., 2015). Advantages- High sensitivity and can be used for large-scale screening. Disadvantages- May produce false positives or negatives and requires specific antibodies.

2.6. Rapid Test Kits

Utilizes antibodies or nucleic acid-based methods for quick detection. Procedure involves various formats like lateral flow assays, immunochromatographic tests, and DNA-based kits. Advantages- Fast, easy to use, and suitable for on-site testing (Posthuma-Trumpie et al., 2009). Disadvantages- Generally, less sensitive than culture-based methods and may require confirmatory tests. These conventional methods are essential tools in microbiology labs for the accurate detection and identification of Salmonella typhimurium. Each method has its own strengths and limitations, and often a combination of methods is used to ensure reliable results.

3. Nanotechnology and Biosensor

Nanotechnology is the science, engineering, and application of materials and devices with structures and properties defined at the nanoscale, which is about 1 to 100 nanometers (nm). These materials exhibit unique physical, chemical, and biological properties that differ significantly from their bulk counterparts, enabling novel applications across various fields, including medicine, electronics, energy, and materials of science. The concept of nanotechnology was first articulated by physicist Richard Feynman in 1959 and later coined by Professor Norio Taniguchi in 1974 (Bayda et al., 2020). Types of nanomaterials include nanoparticles, nanotubes, nanowires, nanofilms and nano coatings, and quantum dots, types and synthesis shown in Figure 2. and Figure 3. Applications of nanotechnology include drug delivery, diagnostics, tissue engineering, electronics, energy, environmental remediation, and materials science. Prospects include nanorobotics, smart materials, and sustainable nanotechnology. Challenges include understanding the long-term effects of nanomaterials on health and the environment, ensuring scalability, and addressing ethical and regulatory issues. Despite these challenges, nanotechnology represents a frontier in scientific and technological advancement. Nanotechnology significantly enhances the performance of biosensors by improving their sensitivity, specificity, and overall performance. Nanomaterials, such as nanoparticles, nanotubes, and nanowires, provide more active sites for biomolecule interaction, leading to increased sensitivity. Quantum effects at the nanoscale enhance the optical, electrical, and magnetic properties of nanomaterials, making biosensors more sensitive to low concentrations of analytes. Electrochemical biosensors and optical biosensors can be amplified by nanomaterials like gold nanoparticles and carbon nanotubes. Nanotechnology also enables the integration of biosensors with microfluidic systems, leading to portable, point-of-care diagnostic devices and lab-on-a-chip platforms. Functionalization and target specificity can be achieved through surface modification and molecule recognition (Kulkarni et al., 2022). Multifunctional nanomaterials can perform multiple functions, enabling versatile biosensors and simultaneous detection of multiple analytes in a single assay. Applications of nanotechnology include medical diagnostics, environmental monitoring, food safety, and agriculture. Recent research examples include gold nanoparticles, carbon nanotubes, and graphene-based biosensors. The relationship between nanotechnology and biosensors is synergistic, allowing for more sensitive, specific, and versatile biosensors (Singh et al., 2026). Biosensors for the rapid detection of Salmonella typhimurium are highly valuable in food safety, clinical diagnostics, and environmental monitoring. Biosensors are analytical devices that combine biological recognition elements with signal transducers to generate measurable outputs.

Figure 2: Classification of nanomaterials

Figure 3: Synthesis of nanomaterials

4. Types of Biosensors

Electrochemical Biosensors: These biosensors detect changes in electrical signals due to the interaction between the sensor and the target bacteria. Components consist of a bioreceptor (like antibodies, aptamers, or enzymes) and a transducer that converts biological interaction into an electrical signal shown in Figure 4. Advantages- High sensitivity, rapid response times, and the potential for miniaturization.

Optical Biosensors: Principle of these biosensors detect changes in optical properties (e.g., fluorescence, absorbance, or luminescence) upon binding with Salmonella typhimurium. Components: They use bioreceptors such as antibodies or DNA probes and an optical transducer. Advantages: High specificity, real-time detection, and the ability to perform multiplexed assays.

Piezoelectric Biosensors: Principle involves these sensors to detect changes in mass or acoustic waves on the sensor surface when Salmonella typhimurium binds to the bioreceptor. Components: Typically, they use quartz crystal microbalances with antibodies or other specific bioreceptors. Advantages: High sensitivity and the ability to detect very low concentrations of bacteria.

Thermal Biosensors: Principle involves these sensors to measure changes in temperature resulting from the metabolic activity of Salmonella typhimurium. Components: They incorporate bioreceptors and a thermal transducer. Advantages: Direct measurement of metabolic activity, which can be highly indicative of bacterial presence 

Figure 4: General schematic of biosensor

5. DNA-Based Biosensor for Salmonella typhimurium:

DNA-based biosensors have emerged as powerful alternatives, offering rapid, selective, and onsite detection of S. typhimurium. These biosensors utilize a DNA probe-often a complementary single-stranded oligonucleotide or aptamer-designed to specifically recognize unique genetic sequences of the pathogen (e.g., invA, fimA, hilA). Integration with nanomaterials and advanced transducers significantly enhances signal transduction, enabling lower detection limits and improved overall assay performance. Because of their portability, high sensitivity, and potential for real-time monitoring, DNA biosensors represent a promising platform for next-generation pathogen detection. This enhanced performance is particularly relevant in pathogen detection, where traditional methods often fall short in terms of speed and accuracy (Virk et al., 2024). The nanocomposite was dropped cast on the glassy carbon electrode and further modified with amino-modified DNA aptamer. The resultant ssDNA/rGO- CNT/GCE aptasensor was then used to detect bacteria by using differential pulse voltammetry (DPV) technique. Synergistic effects of aptasensor were evident through the combination of enhanced electrical properties and facile chemical functionality of both rGO and CNT a consistent nanocomposite interface. Under optimal experimental conditions, the aptasensor could detect S. typhimurium in a wide linear dynamic range from 10¹ until 10⁸ CFU mL⁻¹ with a 10¹ CFU mL⁻¹ of the limit of detection (Appaturi et al.,2020). The electrochemical signal amplification probe was constructed by encapsulating ferrocene into S. typhimurium–specific antimicrobial peptides Magainin I (MI)- Cu₃(PO4)₂ organic-inorganic nanocomposites (Fc@MI) through a one-step process. Magnetic beads (MBs)coupled with antibody were used as a capture ingredient for target magnetic separation, and Fc@MI nanoparticles were used as signal labels in the immunoassays. The sandwich of MBs-target-Fc@MI assay was performed using a screen-printed carbon electrode as a transducer surface. The immunosensor platform presents a low limit of detection (LOD) of 3 CFU mL⁻¹ and a linear range from 10 to 10⁷CFU mL^−1, with good specificity and precision, and was successfully applied for S. typhimurium detection in milk (Bu et al., 2020). The target Salmonella cells were first separated using immunomagnetic nanoparticles and the passive 3D micromixer. Then, immune Au@PtNCs were labeled onto the target cells as signal output to catalyze hydrogen peroxide-3,3′,5,5′- tetramethylbenzidine. Finally, the absorbance was measured at 652 nm to calculate the bacterial amount. This optical biosensor could detect Salmonella at concentrations from 1.8 × 101 to 1.8 × 107 CFU/mL in 1 h. Its detection limit was calculated to be 17 CFU/mL. Besides, this passive 3D micromixer could magnetically separate 99% of target bacteria from the sample in 10 min. This biosensor has the potential to be extended to detect other bacteria by changing the antibodies (L. Zheng et al., 2020). A label-free aptamer was immobilized on a rGOTiO₂ nanocomposite matrix through electrostatic interactions. The changes in electrical conductivity on the electrode surface were evaluated using electroanalytical methods. DNA aptamer adsorbed on the rGO-TiO₂ surface bound to the bacterial cells at the electrode interface causing a physical barrier inhibiting the electron transfer. This interaction decreased the DPV signal of the electrode proportional to decreasing concentrations of the bacterial cells. The optimized aptasensor exhibited high sensitivity with a wide detection range (10⁸ to 10¹ CFU mL⁻¹), a low detection limit of10¹ CFU mL⁻¹ and good selectivity for Salmonella bacteria. This rGO-TiO₂ aptasensor is an excellent biosensing platform that offers a reliable, rapid and sensitive alternative for foodborne pathogen detection (Muniandy et al., 2019). A novel MRS sensor integrated with phages to meet the growing demand for rapid detection of viable Salmonella. A novel phage and CuAAC reaction-based MRS (PCuMRS) biosensor was proposed for rapid, sensitive and cost-effective detection of viable S. typhimurium in food. The conjugates of phage and MNPs with a diameter of 1000 nm (MNP₁₀₀₀-phage), phage and CuO₂@SiO₂-NH₂ nanoparticles (CuO₂@SiO₂-phage), azide (Az) and MNPs with a diameter of 30 nm (MNP₃₀-Az), and alkyne (Alk) and MNP₁₀₀₀ (MNP₁₀₀₀-Alk) were synthesized using the carbodiimide method (Scheme 1A). Thus, established a linear relationship (10²–10⁷ CFU/mL) between the concentration of S. typhimurium and the changes in magnetic signal, with a limit of quantification of 80 CFU/mL (Zhao et al., 2025). A brief review table shown in Table 1.

Dual-Mode Biosensors: A dual-mode biosensor has been developed for the simultaneous and rapid detection of both live and dead Salmonella typhimurium. This biosensor uses bioluminescence and fluorescence detection, providing a rapid and simultaneous assay to distinguish and quantify live and dead bacteria in food samples (Xu et al., 2023). Electrochemical Biosensors: Advances in electrochemical biosensors have improved the detection of foodborne pathogens, including Salmonella. These biosensors utilize CRISPR technology combined with electrochemical detection to enhance sensitivity and specificity, allowing for quick identification of pathogens in food samples (B. Wang et al., 2023). An ultrasensitive ratio metric electrochemical biosensor based on the SRCA-CRISPR/Cas12a system has also been developed. This is the first report of such a biosensor for detecting Salmonella in food, highlighting its high sensitivity and specificity (S. Zheng et al., 2023). Nano-Biosensors: Recent developments include nano-biosensors for the rapid detection of zoonotic bacteria like Salmonella typhimurium. These biosensors leverage nanotechnology to achieve high sensitivity and specificity, making them suitable for various applications, including food safety and environmental monitoring (Ahangari et al., 2023). Lateral flow assays have seen significant advancements for Salmonella detection in food products. These assays often employ bacteriophages engineered to interact specifically with Salmonella cells, providing a simple and rapid detection method suitable for on-site testing (Silva et al., 2023). These recent advancements demonstrate the ongoing efforts to improve the rapid detection of Salmonella typhimurium, making biosensors more efficient, sensitive, and suitable for various applications.

Table 1. Biosensors for detection of Salmonella typhimurium

Abbreviations - Fc@MI – ferrocene functionalized Methylisothiazolinone, COF-AuNPs – covalent organic framework, AuNPs – Gold nanoparticles, MoS₂@Fe₃O_{4 -} magnetite coated molybdenum disulphide, aM – Attomolar.  

6. Challenges and Limitations in Nanomaterial-Based Biosensors

Despite the significant advancements in nanomaterial-enabled biosensors for the detection of Salmonella typhimurium, several challenges still limit their large-scale application and commercialization. These limitations are associated with material properties, device fabrication, operational stability, and real-world applicability shown in Figure 5.

6.1. Reproducibility and Fabrication Issues

One of the major challenges in nanomaterial-based biosensors is the lack of reproducibility during synthesis and device fabrication. Variations in nanomaterial size, shape, and surface chemistry can lead to inconsistent sensor performance. Even slight differences in fabrication conditions can affect sensitivity and signal output. Additionally, uniform immobilization of biomolecules such as DNA probes, antibodies, or aptamers remains difficult to control, further affecting reproducibility (Malik et al., 2023; Patial et al., 2025). Batch-to-batch variation during large-scale production also makes it difficult to standardize biosensor devices for industrial applications.

6.2. Stability and Shelf-Life Limitations

The long-term stability of nanomaterial-based biosensors is another critical concern. Nanomaterials may undergo aggregation, oxidation, or structural degradation over time, which reduces their sensing efficiency. Similarly, biological recognition elements such as enzymes and nucleic acids may lose activity under varying environmental conditions (Pirzada & Altıntaş, 2019; Mishra et al., 2025). Limited shelf-life restricts their practical application, especially in point-of-care and field-based detection systems.

6.3. Matrix Interference in Real Sample Analysis

Although biosensors perform efficiently under laboratory conditions, their performance often decreases in complex real samples such as food and environmental matrices. Interfering substances like proteins, fats, salts, and other microorganisms may cause non-specific interactions and signal interference, leading to inaccurate results (Kabiraz et al., 2023). This remains a major challenge in food safety applications.

6.4. Cost and Scalability Challenges

The cost of nanomaterial synthesis and sensor fabrication remains relatively high due to advanced processing techniques and material requirements. In addition, scaling up biosensor production from laboratory to industrial level without compromising performance is challenging (Kulkarni et al., 2022; Subhan et al., 2025). This limits the commercialization and widespread use of nanomaterial-based biosensors.

Figure 5: Challenges and limitations in nanomaterial-based biosensors

6.5. Functionalization and Surface Modification Complexity

Surface functionalization of nanomaterials is essential for improving specificity and sensitivity. However, achieving stable and uniform functionalization is complex and often involves multiple steps. Improper surface modification can reduce binding efficiency and overall sensor performance (Malik et al., 2023).

6.6. Toxicity and Environmental Concerns

Some nanomaterials, particularly metal-based nanoparticles, may exhibit cytotoxic effects and pose environmental risks. Their accumulation in biological systems and ecosystems can lead to potential health hazards. Therefore, ensuring biocompatibility and environmental safety is important for sustainable biosensor development (Kizhepat et al., 2023).

6.7. Lack of Standardization and Regulatory Approval

The absence of standardized protocols for fabrication, testing, and validation of nano biosensors makes it difficult to compare results across different studies. Furthermore, regulatory approval is limited due to concerns regarding reproducibility, safety, and long-term reliability (Patial et al., 2025).

6.8. Limited Commercialization and Lab-to-Market Gap

Despite extensive research, most nanomaterial-based biosensors remain at the laboratory stage. Challenges such as device integration, cost-effectiveness, and user-friendliness must be addressed for successful commercialization. Bridging the gap between research and industry requires improved scalability and interdisciplinary collaboration (Kulkarni et al., 2022).

7. Recent Advances in Biosensor Development 

7.1. CRISPR-based biosensor: CRISPR-based biosensing systems have recently emerged as powerful tools for the rapid and ultrasensitive detection of foodborne pathogens. The CRISPR-Cas system utilizes programmable guide RNA (crRNA) to recognize specific nucleic acid sequences of pathogens, enabling highly specific detection of bacterial DNA or RNA. When the target sequence is recognized, enzymes such as Cas12 or Cas13 trigger collateral cleavage activity, which can be coupled with nanomaterial-based reporters to generate measurable signals (Wani et al., 2024). In many biosensor platforms, gold nanoparticles (AuNPs) are integrated with CRISPR systems to improve signal amplification and detection sensitivity. For example, CRISPR-Cas12a biosensors combined with AuNP probes can detect the invA gene of Salmonella with extremely high sensitivity. When the target DNA activates Cas12a, the enzyme cleaves single-stranded DNA linkers between AuNP probes, causing nanoparticle dispersion and visible color change. Such platforms have demonstrated detection limits as low as 1 CFU/mL, enabling rapid identification of Salmonella contamination in food samples (Wu et al., 2021). Other nanomaterial-integrated CRISPR biosensors employ silver nanoclusters, magnetic nanoparticles, or nanozymes to enhance signal transduction. Recent systems such as the SCENT-Cas platform combine CRISPR-Cas12a with fluorescent silver nanoclusters to detect Salmonella typhimurium with high sensitivity and a dynamic detection range from 1 to 10⁸ CFU/mL (Wani et al., 2024). These CRISPR-nano biosensors offer several advantages, including rapid detection, programmable specificity, minimal sample preparation, and compatibility with portable detection platforms. Consequently, they represent a promising approach for next generation food safety monitoring systems.

7.2. Smartphone-Integrated Biosensors: Recent advances in portable diagnostics have led to the development of smartphone-integrated biosensors for pathogen detection. These systems combine nanomaterial-based biosensors with smartphone cameras, microfluidic chips, or optical detectors to provide rapid and on-site analysis. Smartphones can capture fluorescence, colorimetric, or electrochemical signals generated by biosensors and convert them into quantitative results using dedicated mobile applications. In several studies, CRISPR-Cas12-based detection platforms integrated with smartphones have demonstrated ultrasensitive detection of foodborne pathogens with limits of detection as low as 1 CFU/mL (Xie et al., 2024). In addition to portability, smartphone-based biosensors can incorporate artificial intelligence (AI) algorithms for automated signal analysis, image processing, and pattern recognition. AI-assisted analysis improves accuracy, reduces human error, and enables rapid data interpretation in field conditions. These technologies are particularly useful for food safety monitoring in remote areas, agricultural environments, and supply chain inspection systems.

7.3. Paper-Based and Microfluidic Devices: Paper-based biosensors and microfluidic devices have attracted significant attention due to their low cost, portability, and ease of use. These platforms typically use cellulose paper or polymer microchannels to guide sample flow and enable biochemical reactions within a miniaturized system. Microfluidic paper-based analytical devices (µPADs) integrated with nanomaterials and nucleic acid amplification techniques can rapidly detect pathogens in food samples. For instance, a microfluidic paper device combined with recombinase polymerase amplification (RPA) and surface-enhanced Raman spectroscopy (SERS) has been developed for rapid detection of Salmonella typhimurium, achieving detection within 45 minutes with high sensitivity and specificity (Wani et al., 2024). Because of these features, paper-based biosensors are particularly suitable for point-of-care (POC) and field-based pathogen detection in food safety applications.

7.4. Multiplex Detection Platforms: Another significant advancement in biosensor technology is the development of multiplex detection systems capable of simultaneously identifying multiple pathogens in a single assay. Multiplex biosensors are highly desirable in food safety monitoring because food samples often contain several microbial contaminants. Nanomaterial-based signal tags such as quantum dots, fluorescent nanoparticles, and magnetic nanocomposites enable simultaneous detection of multiple targets by generating distinct optical or electrochemical signals. CRISPR-based multiplex biosensors have also been developed using different Cas enzymes or guide RNAs to detect multiple bacterial species simultaneously (Sun et al., 2024). For example, dual-CRISPR fluorescence biosensors can detect multiple pathogens by using different fluorescent probes that produce distinguishable signals. Such systems allow simultaneous detection of pathogens like Staphylococcus aureus and Pseudomonas aeruginosa in a single assay, general structure shown in Figure 6.

Figure 6: Recent advancements in biosensor development

8. Conclusion

The growing global burden of infectious and foodborne diseases highlights an urgent need for rapid, accurate, and reliable diagnostic tools. Biosensors, enhanced by advances in nanotechnology, have emerged as powerful alternatives to conventional detection methods, offering higher sensitivity, faster response times, and greater specificity. Nanomaterials such as graphene, metal oxides, carbon nanotubes, and quantum dots provide unique physicochemical advantages that significantly improve biosensor performance across healthcare, environmental monitoring, food safety, and biodefense applications. This review emphasizes the critical role of biosensors in detecting pathogens like Salmonella typhimurium, where early identification is essential for preventing outbreaks and reducing public health risks. While culture-based, biochemical, immunological, and molecular techniques remain foundational, nanomaterial-enabled biosensors demonstrate superior efficiency and adaptability for real-time detection. Recent advancements in electrochemical, optical, piezoelectric, thermal, and DNA-based biosensors showcase the rapid progress being made toward highly sensitive, portable, and field-deployable diagnostic systems. Overall, the integration of nanotechnology with biosensor platforms holds immense promise for transforming disease detection, improving public health, and enabling more responsive and accessible diagnostic systems in the future.

9. Prospects

It is anticipated that the development of biosensors for infectious diseases would transform approaches to illness management, prevention, and detection. Emerging technologies and platforms, tailored therapy and management, real-time monitoring and surveillance, early detection and diagnosis, and these are some of the main areas of concentration. The ability to identify infectious agents at low concentrations in a variety of sample types will be made possible by high sensitivity and specificity. Comprehensive diagnostic testing will be made easier with the use of multiplexed detection, which enables the simultaneous identification of multiple infections or biomarkers linked to various diseases. Continuous monitoring of illness progression, response to treatment, and possible outbreaks will be made possible via real-time monitoring. Additionally, biosensors will speed up the identification of antimicrobial resistance indicators, assisting in the formulation of suitable treatment plans. Targeted antimicrobial agent distribution to certain infection locations will be made possible by integrated therapies. upcoming expenditures on cooperation, innovation, and research. Also, till now biosensor development is mostly restricted to laboratories. A consistent, cost-effective approach is required to expand biosensor to market level. As for the use of common people, biosensors are highly useful in detecting bioterrorism, where long diagnosis procedures are not practical.

CRediT authorship contribution statement

Shivangi Kosma: Conceptualization, Data Curation, Methodology, Writing-Original Draft,

Riya Ritika Singh: Visualization, Methodology, Review and Editing.

Manoj Kumar Patel: Visualization, Supervision, Review and Editing.  

Statement for Use of Generative AI and Tools

Quill Bot has been used for Paraphrasing only. Writing, review insights, and data are done by authors.

Declaration of competing interest

There is no conflict of interest regarding the publication of this paper. 

Acknowledgment

RRS is thankful to the Department of Science and Technology (DST), New Delhi, India for the award of INSPIRE fellowship (No. DST/INSPIRE Fellowship/2020/IF200007). And MKP acknowledges Pt. Ravishankar Shukla University, Raipur for Seed Money Research Grant (Letter No. 2364/Acad./RGSM/2025).   

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