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Updated: Aug 9, 2026
Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria
Published on: May 8, 2013
Fluorometric Paper-Based, Loop-Mediated Isothermal Amplification Devices for Quantitative Point-of-Care Detection of
Ilada Choopara1, Akkapol Suea-Ngam2, Yothin Teethaisong3
1Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
Abstract:
Loop-mediated isothermal amplification (LAMP) has been widely used to detect many infectious diseases. However, minor inconveniences during the steps of adding reaction ingredients and lack of simple color results hinder point-of-care detection. We therefore invented a fluorometric paper-based LAMP by incorporating LAMP reagents, including a biotinylated primer, onto a cellulose membrane paper, with a simple DNA fluorescent dye incubation that demonstrated rapid and accurate results parallel to quantitative polymerase chain reaction (qPCR) methods. This technology allows for instant paper strip detection of methicillin-resistant Staphylococcus aureus (MRSA) in the laboratory and clinical samples. MRSA represents a major public health problem as it can cause infections in different parts of the human body and yet is resistant to commonly used antibiotics. In this study, we optimized LAMP reaction ingredients and incubation conditions following a central composite design (CCD) that yielded the shortest reaction time with high sensitivity. These CCD components and conditions were used to construct the paper-based LAMP reaction by immobilizing the biotinylated primer and the rest of the LAMP reagents to produce the ready-to-use MRSA diagnostic device. Our paper-based LAMP device could detect as low as 10 ag (equivalent to 1 copy) of the MRSA gene mecA within 36-43 min, was evaluated using both laboratory (individual cultures of MRSA and non-MRSA bacteria) and clinical blood samples to be 100% specific and sensitive compared to qPCR results, and had 35 day stability under 25 °C storage. Furthermore, the color readout allows for quantitation of MRSA copies. Hence, this device is applicable for point-of-care MRSA detection.
Insights
A new paper-based assay simplifies infectious disease detection using loop-mediated isothermal amplification (LAMP). This rapid, sensitive diagnostic tool accurately identifies methicillin-resistant Staphylococcus aureus (MRSA) for point-of-care use.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Point-of-Care Testing
Background:
- Loop-mediated isothermal amplification (LAMP) is vital for infectious disease detection but faces practical limitations for point-of-care applications.
- Current methods often involve complex reagent handling and lack simple, visual results.
- Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant public health threat due to antibiotic resistance.
Purpose of the Study:
- To develop a user-friendly, paper-based fluorometric LAMP assay for rapid MRSA detection.
- To overcome the limitations of traditional LAMP assays for point-of-care settings.
- To create a sensitive and specific diagnostic tool for MRSA identification.
Main Methods:
- Incorporation of LAMP reagents and a biotinylated primer onto a cellulose membrane paper.
- Optimization of LAMP reaction conditions using a central composite design (CCD) for speed and sensitivity.
- Development of a simple DNA fluorescent dye incubation for result readout.
Main Results:
- The paper-based LAMP assay detected as low as 10 attograms (1 copy) of the MRSA mecA gene within 36-43 minutes.
- The assay demonstrated 100% sensitivity and specificity compared to quantitative polymerase chain reaction (qPCR) using laboratory and clinical samples.
- The device exhibited 35-day stability at 25 °C and allowed for colorimetric quantitation of MRSA copies.
Conclusions:
- The developed fluorometric paper-based LAMP assay is a rapid, sensitive, and specific tool for MRSA detection.
- This ready-to-use diagnostic device is suitable for point-of-care applications, addressing limitations of existing methods.
- The technology offers a promising solution for managing MRSA infections in diverse healthcare settings.
