Food safety is a major issue because of the prevalence of pathogens, chemicals, and other potentially harmful substances in food products. Because of this, the creation of a reliable and accessible tool for checking on food quality is essential. In comparison to traditional techniques, lateral flow tests for the identification of food-borne pathogens are more modern, straightforward, and quick. This review examines the use of lateral flow biosensors for the detection of various food contaminants and the labels used to improve the effectiveness of the system. Since LFAs may be carried out by non-specialists, they are well-suited for on-site testing in a variety of food production and distribution environments. 

Lateral flow assays for food analyses: Food contaminants, allergens, toxins, and beyond

The COVID-19 pandemic has spurred demand for efficient and rapid diagnostic tools that can be deployed at point of care to quickly identify infected individuals. Existing detection methods are time consuming and they lack sensitivity. Point-of-care testing (POCT) has emerged as a promising alternative due to its user-friendliness, rapidity, and high specificity and sensitivity. Such tests can be conveniently conducted at the patient’s bedside. Immunodiagnostic methods that offer the rapid identification of positive cases are urgently required. Quantum dots (QDs), known for their multimodal properties, have shown potential in terms of combating or inhibiting the COVID-19 virus. When coupled with specific antibodies, QDs enable the highly sensitive detection of viral antigens in patient samples. Conventional lateral flow immunoassays (LFAs) have been widely used for diagnostic testing due to their simplicity, low cost, and portability. However, they often lack the sensitivity required to accurately detect low viral loads. Quantum dot (QD)-based lateral flow immunoassays have emerged as a promising alternative, offering significant advancements in sensitivity and specificity. Moreover, the lateral flow immunoassay (LFIA) method, which fulfils POCT standards, has gained popularity in diagnosing COVID-19. This review focuses on recent advancements in QD-based LFIA for rapid POCT COVID-19 diagnosis. Strategies to enhance sensitivity using QDs are explored, and the underlying principles of LFIA are elucidated. The benefits of using the QD-based LFIA as a POCT method are highlighted, and its published performance in COVID-19 diagnostics is examined. Overall, the integration of quantum dots with LFIA holds immense promise in terms of revolutionizing COVID-19 detection, treatment, and prevention, offering a convenient and effective approach to combat the pandemic. 

Recent Advances in Quantum Dot-Based Lateral Flow Immunoassays for the Rapid, Point-of-Care Diagnosis of COVID-19

Khapra beetle (Trogoderma granarium Everts), one of the most important quarantine pests globally, is capable of causing severe infestation and huge economic loss to stored grain, and its interception rate has increased in major global trade countries over the past few years. However, difficulties remain in distinguishing this species with similar ones. In order to assist border ports and warehouses in khapra beetle's effective rapid identification as well as pest control at the early stages of monitoring or interception, we herein developed a new and rapid visual detection assay for T. granarium based on recombinase polymerase amplification (RPA) and the CRISPR/Cas12a system.We designed and selected the first khapra beetle-specific RPA primers and crRNA, and optimized the visualization reaction system (Cas12a/CrRNA = 100 nM/500 nM). With only a 37 °C-heat-source and a blue light torch, RPA and CRISPR/CAS12a-based visualization assays can be completed within 40 min to differentiate between khapra beetle and nine similar Dermestidae species. After DNA extraction using a kit (4-5 h) or a simple method (5 min), the specific amplicons were obtained after a 15 min RPA reaction at 37 °C, followed by a 15 min color reaction under 37 °C in dark conditions using a CRISPR/CAS12a system and a fluorescent probe (5'-FAM/3'-BHQ1 labeled). This method is ingenious to low levels of DNA (10-1 ng μL-1 ) and meets the sensitivity requirements for detecting a single khapra beetle's egg (≈0.7 mm).Our specificity and sensitivity analysis inferred that the present visualization system is effective to quickly and uniquely detect khapra beetle at room temperature (37 °C), thereby preventing this species before they spread widely. Our study is suitable for being pushed forward in storage pest management, and provides value as a reference for monitoring and identification of other pests. © 2023 Society of Chemical Industry. 

New and rapid visual detection assay for Trogoderma granarium Everts based on recombinase polymerase amplification and CRISPR/Cas12a.

Lateral flow immunoassays (LFIAs) serve as rapid and convenient tools for disease screening. However, there is room for enhancing diagnostic sensitivity. Therefore, this paper, introduces a novel material combined with lateral flow immunoassays to improve the sensitivity of COVID-19 detection and diagnosis. Trimethylsilyl cellulose (TMSC) was printed on the device between the test and control line to delay the flow of the solution, which resulted in a more excellent binding of the immunoassay. These delay-LFIAs (d-LFIAs) were developed for the diagnosis of COVID-19, targeting the detection of immunoglobulins G and M, as well as the COVID-19 antigen. Under optimized conditions, the d-LFIA increased the sensitivity by up to 5 times compared to that of conventional LFIAs. Moreover, it exhibited the ability to detect IgG, IgM, and SARS-CoV-2 antigen as low as 1 ng/mL (limit of detection) by the naked eye. In addition, the application of this device to real samples yielded highly consistent results with those obtained from other LFIAs and standard SARS-CoV-2 diagnostic methods (such as enzyme-linked immunosorbent assay and reverse transcription polymerase chain reaction) diagnosis. Therefore, we expect this device to be new platform for highly sensitive, easy, and affordable immunoassay strip testing to certify future emerging diseases. 

Integrated lateral flow immunoassays using trimethylsilyl cellulose barriers for the enhanced sensitivity of COVID-19 diagnosis

Staphylococcus aureus is a species of Staphylococcus and belongs to Gram-positive. Meanwhile, S. aureus remains one of the most common nosocomial and community-acquired infections, causing blood flow, skin, soft tissue, and lower respiratory tract infections. ABSTRACT Antimicrobial resistance (AMR) has posed a global threat to public health. The Staphylococcus aureus strains have especially developed AMR to practically all antimicrobial medications. There is an unmet need for rapid and accurate detection of the S. aureus AMR. In this study, we developed two versions of recombinase polymerase amplification (RPA), the fluorescent signal monitoring and lateral flow dipstick, for detecting the clinically relevant AMR genes retained by S. aureus isolates and simultaneously identifying such isolates at the species level. The sensitivity and specificity were validated with clinical samples. Our results showed that this RPA tool was able to detect antibiotic resistance for all the 54 collected S. aureus isolates with high sensitivity, specificity, and accuracy (all higher than 92%). Moreover, results of the RPA tool are 100% consistent with that of PCR. In sum, we successfully developed a rapid and accurate AMR diagnostic platform for S. aureus. The RPA might be used as an effective diagnostic test in clinical microbiology laboratories to improve the design and application of antibiotic therapy. IMPORTANCE Staphylococcus aureus is a species of Staphylococcus and belongs to Gram-positive. Meanwhile, S. aureus remains one of the most common nosocomial and community-acquired infections, causing blood flow, skin, soft tissue, and lower respiratory tract infections. The identification of the particular nuc gene and the other eight genes of drug-resistant S. aureus can reliably and quickly diagnose the illness, allowing doctors to prescribe treatment regimens sooner. The detection target in this work is a particular gene of S. aureus, and a POCT is built to simultaneously recognize S. aureus and analyze genes representing four common antibiotic families. We developed and assessed a rapid and on-site diagnostic platform for the specific and sensitive detection of S. aureus. This method allows the determination of S. aureus infection and 10 different AMR genes representing four different families of antibiotics within 40 min. It was easily adaptable in low-resource circumstances and professional-lacking circumstances. It should be supported in overcoming the continuous difficulty of drug-resistant S. aureus infections, which is a shortage of diagnostic tools that can swiftly detect infectious bacteria and numerous antibiotic resistance indicators.

https://doi.org/10.1128/spectrum.04476-22

A Rapid Antimicrobial Resistance Diagnostic Platform for Staphylococcus aureus Using Recombinase Polymerase Amplification

ABSTRACT Lateral flow immunoassay is the leading Point of Care test and is becoming increasingly essential for its versatile properties. The attraction of lateral flow assay (LFA) has reached its prime position during recent SARS-CoV-2 pandemic and Ebola, Zika epidemics in third world countries where primary screening of the disease and financial issues are very important. During the last decade traditional methodology of LFA was limited to visual detection and qualitative assessment only. However, recently researchers are focusing on the development and improvement of this tool to enhance its specificity, assessment power (quantitative) to make it an alternative to traditional lab-based technology. Modifying working principle and instrumentation, combination of different modern molecular techniques such as Reverse transcription loop mediated isothermal amplification (RT-LAMP), Clustered regularly inter-spaced short palindromic repeat (CRISPR-Cas), Recombinase amplification polymerase (RPA), also association of image-based software, involvement of nanotechnology, implementation of LFA ruler have established authenticity and ultra-specific detection level. These leading immunochromatographic techniques offer simultaneous detection of different analytes from a single sample unit into one multiplex strip and provide the necessary information. This review is a foremost attempt to encompass recent advances of lateral flow assays in combination with molecular biology techniques along with improvements of assay components for improved diagnostic sensitivity and specificity. Some infectious disease diagnosis by LFA with its reporter and low detection limit have also been mentioned in this review.

Recent advances of lateral flow immunoassay components as “point of need”

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Recent advances of lateral flow immunoassay components as “point of need”