Sensitive colorimetric detection of antibiotic resistant Staphylococcus aureus on dairy farms using LAMP with pH-responsive polydiacetylene

Li, Qiaofeng; An, Zhaoxia; Sun, Tieqiang; Ji, Shuaifeng; Wang, Weiya; Peng, Yuan; Wang, Zhouping; Salentijn, Gert Ij; Gao, Zhixian; Han, Dianpeng

doi:10.1016/j.bios.2022.114824

Cited by 6 publications

(3 citation statements)

References 45 publications

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“…By eliminating complex thermal cycling processes, these techniques can simplify the operation and reduce detection time. Isothermal amplification methods such as loop-mediated isothermal amplification (LAMP) [ 11 , 12 ], rolling circle amplification (RCA) [ 13 , 14 ], strand displacement amplification (SDA) [ 15 , 16 ], and recombinase polymerase amplification (RPA) [ 17 , 18 ] offer the benefits of low cost, high sensitivity, and high specificity. Of these, RPA has demonstrated significant potential for practical applications due to its capacity to perform rapid detection in under 40 min at a consistent temperature of 37–40 °C, by introducing recombinase, strand-substitution polymerase, and ssDNA-binding protein (SSB) [ 19 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

An Integrated Microfluidic Biosensing System Based on a Versatile Valve and Recombinase Polymerase Amplification for Rapid and Sensitive Detection of Salmonella typhimurium

Jin

Wang

et al. 2023

Biosensors

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Detecting foodborne pathogens on-site is crucial for ensuring food safety, necessitating the development of rapid, cost-effective, highly sensitive, and portable devices. This paper presents an integrated microfluidic biosensing system designed for the rapid and sensitive detection of Salmonella typhimurium (S. typhimurium). The biosensing system comprises a microfluidic chip with a versatile valve, a recombinase polymerase amplification (RPA) for nucleic acid detection, and a customized real-time fluorescence detection system. The versatile valve combines the functions of an active valve and a magnetic actuation mixer, enabling on-demand mixing and controlling fluid flow. Quantitative fluorescence is processed and detected through a custom-built smartphone application. The proposed integrated microfluidic biosensing system could detect Salmonella at concentrations as low as 1.0 × 102 copies/µL within 30 min, which was consistent with the results obtained from the real-time quantitative polymerase chain reaction (qPCR) tests. With its versatile valve, this integrated microfluidic biosensing system holds significant potential for on-site detection of foodborne pathogens.

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Section: Introductionmentioning

confidence: 99%

An Integrated Microfluidic Biosensing System Based on a Versatile Valve and Recombinase Polymerase Amplification for Rapid and Sensitive Detection of Salmonella typhimurium

Jin

Wang

et al. 2023

Biosensors

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“…Emerging methodologies based on nucleic acids appear to be superior to other methods since they are capable of directly identifying the related antibiotic resistance genes. The sensitivity of established and developing isothermal amplification techniques, such as loop-mediated isothermal amplification and nucleic acid sequence-based amplification, is commensurate with that of the general polymerase chain reaction (PCR) [ 11 - 14 ]. For instance, Luyu Wei et al .…”

Section: Introductionmentioning

confidence: 99%

Sensitive and Extraction-Free Detection of Methicillin-Resistant Staphylococcus aureus through Ag⁺ Aptamer-Based Color Reaction

Cao,

Zhang,

et al. 2023

J. Microbiol. Biotechnol.

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Refractory infections, such as hospital-acquired pneumonia, can be better diagnosed with the assistance of precise methicillin-resistant Staphylococcus aureus (MRSA) testing. However, traditional methods necessitate high-tech tools, rigorous temperature cycling, and the extraction of genetic material from MRSA cells. Herein, we propose a sensitive, specific, and extraction-free strategy for MRSA detection by integrating allosteric probe-based target recognition and exonuclease-III (Exo-III)-enhanced color reaction. The penicillin-binding protein 2a (PBP2a) aptamer in the allosteric probe binds with MRSA to convert protein signals to nucleic acid signals. This is followed by the DNA polymerase-assisted target recycle and the production of numerous single-strand DNA (ssDNA) chains which bind with silver ion (Ag + ) aptamer to form a blunt terminus that can be identified by Exo-III. As a result, the Ag + aptamer pre-coupled to magnetic nanoparticles is digested. After magnetic separation, the Ag + in liquid supernatant catalyzes 3,3’,5,5’-tetramethylbenzidine (TMB) for a color reaction. In addition, a concentration of 54 cfu/mL is predicted to be the lowest detectable value. Based on this, our assay has a wide linear detection range, covering 5 orders of magnitude and demonstrating a high specificity, which allows it to accurately distinguish the target MRSA from other microorganisms.

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“…For DNA amplification, isothermal techniques have recently gathered much attention from researchers owing to their rapidity, cost effectiveness, ease of use, and ability to be performed under constant temperature. Some common isothermal techniques include loop-mediated isothermal amplification (LAMP) [13][14][15], rolling circle amplification (RCA) [16,17], nucleic acid sequence-based amplification (NASBA) [18,19], and recombinase polymerase amplification (RPA) [20,21]. Among these listed isothermal techniques, LAMP serves as the most promising approach because it requires less enzyme compared to RPA.…”

Section: Introductionmentioning

confidence: 99%

Pipette-Free and Fully Integrated Paper Device Employing DNA Extraction, Isothermal Amplification, and Carmoisine-Based Colorimetric Detection for Determining Infectious Pathogens

Nguyen,

Lee

2023

Sensors

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A pipette-free and fully integrated device that can be used to accurately recognize the presence of infectious pathogens is an important and useful tool in point-of-care testing, particularly when aiming to decrease the unpredictable threats posed by disease outbreak. In this study, a paper device is developed to integrate the three main processes required for detecting infectious pathogens, including DNA extraction, loop-mediated isothermal amplification (LAMP), and detection. All key reagents, including sodium dodecyl sulfate (SDS), NaOH, LAMP reagents, and carmoisine, are placed on the paper device. The paper device is operated simply via sliding and folding without using any bulky equipment, and the results can be directly observed by the naked eye. The optimized concentrations of sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), and carmoisine were found to be 0.1%, 0.1 M, and 0.5 mg/mL, respectively. The paper device was used to detect Enterococcus faecium at concentrations as low as 102 CFU/mL within 60 min. Also, E. faecium spiked in milk was successfully detected using the paper device, demonstrating the feasible application in real sample analysis.

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Sensitive colorimetric detection of antibiotic resistant Staphylococcus aureus on dairy farms using LAMP with pH-responsive polydiacetylene

Cited by 6 publications

References 45 publications

An Integrated Microfluidic Biosensing System Based on a Versatile Valve and Recombinase Polymerase Amplification for Rapid and Sensitive Detection of Salmonella typhimurium

An Integrated Microfluidic Biosensing System Based on a Versatile Valve and Recombinase Polymerase Amplification for Rapid and Sensitive Detection of Salmonella typhimurium

Sensitive and Extraction-Free Detection of Methicillin-Resistant Staphylococcus aureus through Ag⁺ Aptamer-Based Color Reaction

Pipette-Free and Fully Integrated Paper Device Employing DNA Extraction, Isothermal Amplification, and Carmoisine-Based Colorimetric Detection for Determining Infectious Pathogens

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Sensitive colorimetric detection of antibiotic resistant Staphylococcus aureus on dairy farms using LAMP with pH-responsive polydiacetylene

Cited by 6 publications

References 45 publications

An Integrated Microfluidic Biosensing System Based on a Versatile Valve and Recombinase Polymerase Amplification for Rapid and Sensitive Detection of Salmonella typhimurium

An Integrated Microfluidic Biosensing System Based on a Versatile Valve and Recombinase Polymerase Amplification for Rapid and Sensitive Detection of Salmonella typhimurium

Sensitive and Extraction-Free Detection of Methicillin-Resistant Staphylococcus aureus through Ag+ Aptamer-Based Color Reaction

Pipette-Free and Fully Integrated Paper Device Employing DNA Extraction, Isothermal Amplification, and Carmoisine-Based Colorimetric Detection for Determining Infectious Pathogens

Contact Info

Product

Resources

About

Sensitive and Extraction-Free Detection of Methicillin-Resistant Staphylococcus aureus through Ag⁺ Aptamer-Based Color Reaction