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Updated: Jan 31, 2026
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Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System
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Rapid and Sensitive Pathogen Detection and Viability Evaluation: A Lateral Flow Assay Based on One-to-Multiple Signal
Yang Liu1,2, Qingfeng Zheng1, Tao Liao3
1Institute of Biotechnology, Xianghu Laboratory, Hangzhou 311231, China.
Analytical Chemistry
|January 29, 2026
Summary
This study introduces NALFIA, a novel assay improving pathogen detection sensitivity and efficiency. It differentiates live and dead pathogens, offering a cost-effective solution for point-of-care diagnostics and antibacterial assessments.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Assay Development
Background:
- Lateral flow assays (LFAs) are crucial for point-of-care pathogen detection.
- Existing LFAs face limitations in sensitivity and pathogen viability differentiation.
Purpose of the Study:
- To develop a novel assay, NALFIA, enhancing sensitivity and viability detection for pathogens.
- To improve upon existing lateral flow assay technologies for diagnostics.
Main Methods:
- Integration of DNA-binding molecules, asymmetric isothermal amplification, and immuno-LFA.
- Utilized a one-to-multiple secondary signal amplification strategy with single-stranded amplicons, F-probes, and PSred nanospheres.
- Validated NALFIA with 90 real samples against quantitative polymerase chain reaction (qPCR).
Main Results:
- NALFIA achieved approximately 100-fold improved detection limit compared to single-probe LFAs.
- Results were obtainable by naked eye within 5 minutes, demonstrating high efficiency.
- 100% concordance with qPCR was observed in real-sample validation.
- Demonstrated capability to assess pathogen viability, shown in UV-induced germicidal efficacy studies.
Conclusions:
- NALFIA offers a highly sensitive, cost-effective, and rapid platform for pathogen detection.
- The assay can distinguish between live and dead pathogens, crucial for efficacy assessments.
- NALFIA is suitable for mass pathogen screening and field-deployable applications.
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