Loop-Mediated Isothermal Amplification Coupled With Nanoparticle-Based Biosensor: A Rapid and Sensitive Method to...

Area of Science:

• Microbiology
• Molecular Diagnostics
• Biotechnology

Background:

• Mycoplasma pneumoniae (MP) causes MP pneumonia (MPP), a significant public health concern.
• Current diagnostic methods for MP lack speed and efficiency, hindering timely treatment.
• There is a critical need for rapid, sensitive, and specific diagnostic tools for MP detection.

Purpose of the Study:

• To develop a novel loop-mediated isothermal amplification (LAMP)-based assay coupled with a nanoparticle-based lateral flow biosensor (LAMP-LFB).
• To achieve rapid, sensitive, and specific diagnosis of Mycoplasma pneumoniae (MP) infections.
• To validate the assay's performance using clinical samples and compare it with existing methods.

Main Methods:

• A set of six primers targeting the community-acquired respiratory distress syndrome (CARDS) toxin gene of MP was designed.
• The LAMP assay was optimized to run at 63°C for 30 minutes.
• The LAMP products were detected using a lateral flow biosensor (LFB) within 2 minutes, enabling a total assay time of under one hour.

Main Results:

• The MP-LAMP-LFB assay demonstrated high sensitivity with a detection limit of 50 fg per reaction.
• The assay showed no cross-reactivity with other common respiratory pathogens.
• Validation with 100 nasopharyngeal swab samples from children suspected of MPP revealed a 50% positive rate, consistent with real-time PCR results.

Conclusions:

• The developed MP-LAMP-LFB assay is a rapid, highly sensitive, and specific diagnostic tool for Mycoplasma pneumoniae.
• This assay is suitable for point-of-care settings and basic medical facilities, especially in rural areas.
• The MP-LAMP-LFB assay offers a valuable alternative for timely MP diagnosis and management.