A 3D-Printed Integrated Handheld Biosensor for the Detection of Vibrio parahaemolyticus

Area of Science:

• Food Safety
• Biosensor Technology
• Molecular Diagnostics

Background:

• Vibrio parahaemolyticus is a significant seafood-borne pathogen causing human gastrointestinal illness.
• Biosensors offer precise detection of foodborne pathogens, but low-cost, portable devices are needed.
• Current detection methods can be time-consuming and require specialized equipment.

Purpose of the Study:

• To develop a low-cost, integrated handheld biosensor (IHB) for ultrasensitive detection of Vibrio parahaemolyticus.
• To combine Recombinase Polymerase Amplification (RPA), CRISPR/Cas12a, and Lateral Flow Strips (LFS) in a portable device.
• To enable rapid and accurate screening of Vibrio parahaemolyticus in food samples.

Main Methods:

• Developed a 3D-printed IHB integrating RPA, CRISPR/Cas12a, and LFS.
• Utilized RPA to preamplify the tlh gene of Vibrio parahaemolyticus.
• Employed CRISPR/Cas12a trans-cleavage activity to detect the amplified DNA via LFS.

Main Results:

• The IHB demonstrated high selectivity and excellent sensitivity for Vibrio parahaemolyticus detection.
• Achieved an ultrasensitive limit of detection of 4.9 CFU/mL.
• The system showed a negative correlation between amplified RPA products and LFS signal intensity.

Conclusions:

• The developed IHB is a promising tool for the ultrasensitive and rapid screening of Vibrio parahaemolyticus in samples.
• This technology has the potential for broader application in monitoring seafood and marine environmental safety for various pathogen risks.
• The low-cost, portable nature of the IHB facilitates widespread use in food safety applications.