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Biotinylation-based lateral flow assays for pathogenic and total bacteria detection.

Yongqiang Cao¹, Yanlin Chen¹, Xinyi Zhang¹

¹Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education, Guangxi Colleges and Universities Key Laboratory of Biological Molecular Medicine Research, School of Basic Medical Sciences, Guangxi Medical University, Nanning, Guangxi, 530021, PR China.

Analytica Chimica Acta

|January 20, 2025

Summary

A novel biotinylation-based lateral flow assay (LFA) enables simultaneous detection of total and pathogenic bacteria. This advancement overcomes limitations in traditional LFAs, offering rapid bacterial detection for water quality management.

Keywords:

Biotinylation Detection Lateral flow assay Pathogenic Total bacteria

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Area of Science:

Microbiology
Biotechnology
Analytical Chemistry

Background:

Current lateral flow assays (LFAs) are effective for detecting pathogenic bacteria but struggle with total bacteria detection due to the absence of a universal antibody.
Concurrent evaluation of both pathogenic and total bacteria is crucial for water and food safety monitoring.

Purpose of the Study:

To develop a novel biotinylation-based lateral flow assay (LFA) capable of detecting both total and pathogenic bacteria.
To overcome the limitations of traditional LFAs in identifying the overall bacterial load.

Main Methods:

Nonspecific labeling of bacterial surface lipoproteins using sulfo-NHS-biotin.
Preparation of streptavidin-coated gold nanoparticles (Str-AuNP) and test lines.
Integration of biotinylation into the LFA platform for universal bacterial recognition.

Main Results:

A biotinylation-based LFA was successfully established for the detection and semi-quantification of total bacteria.
The assay demonstrated good practicability in detecting total bacteria in real-world water samples (pond and wastewater).
The platform was also adapted for pathogenic bacteria detection using specific biotinylation, without requiring new test strip preparation.

Conclusions:

The integration of nonspecific bacterial biotinylation into LFAs is a novel approach to address total bacteria detection challenges.
The developed biotinylation-based LFAs offer a versatile tool for detecting both total and pathogenic bacteria simultaneously.
This method holds significant potential for rapid bacterial detection in water quality management and control.