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Quantitative PCR of T7 Bacteriophage from Biopanning

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Rapid Bacteriophage Quantification by Digital Biosensing on a SlipChip Microfluidic Device.

Xiang Li¹, Qixin Hu¹, Xu Liu¹

¹School of Biomedical Engineering, Shanghai Jiao Tong University, 1954 Hua Shan Road, Shanghai 200030, China.

Analytical Chemistry

|May 22, 2023

Summary

A new digital biosensing method rapidly quantifies infectious bacteriophages using a microfluidic device. This advancement aids phage therapy development for combating antimicrobial resistance (AMR) and offers a sensitive bacterial detection tool.

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Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria

Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria

Published on: May 8, 2013

Area of Science:

Microbiology
Biotechnology
Bioengineering

Background:

Antimicrobial resistance (AMR) is a critical global health threat, necessitating novel therapeutic strategies.
Bacteriophage therapy is a promising alternative for treating AMR, but accurate phage quantification is essential for its clinical application.
Existing phage quantification methods are time-consuming, labor-intensive, or unable to distinguish infectious from non-infectious phages.

Purpose of the Study:

To develop a rapid, precise, and reliable digital biosensing method for quantifying infectious bacteriophages.
To establish a microfluidic platform for high-throughput phage enumeration.
To provide a tool that supports the clinical development and application of phage therapy.

Main Methods:

Development of a digital phage SlipChip (dp-SlipChip) microfluidic device with 2304 nanoliter microdroplets.
Compartmentalization of bacteriophages and bacteria within microdroplets for simultaneous analysis.
Quantification of infectious phages based on bacterial growth profiles within 3 hours.

Main Results:

The dp-SlipChip accurately quantified infectious phages, demonstrating results consistent with traditional plaque assays.
The digital biosensing method exhibited superior consistency and repeatability compared to conventional techniques.
The system achieved rapid quantification (3 hours) without requiring complex fluidic handling instruments.

Conclusions:

The dp-SlipChip offers a significant advancement for rapid and precise bacteriophage quantification, crucial for effective phage therapy against AMR.
This digital biosensing approach provides an ultrasensitive and highly specific method for bacterial detection.
The technology holds potential for broader applications in digital biology studies requiring single-object analysis.