Development of multisample detection system using a tag insertion primer and an electrochemical DNA chip

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Bioanalytical Chemistry

Background:

• Accurate and efficient detection of infectious agents is crucial for public health.
• Existing diagnostic methods, such as nested polymerase chain reaction (PCR), can be time-consuming and costly.
• There is a need for high-throughput, cost-effective diagnostic tools.

Purpose of the Study:

• To develop and validate a novel multisample detection system.
• To combine tag insertion primer technology with an electrochemical DNA chip for enhanced detection.
• To assess the system's performance in detecting Helicobacter species infections.

Main Methods:

• Utilized tag insertion primers to introduce unique tag sequences into amplification products from each sample.
• Employed loop-mediated isothermal amplification (LAMP) to generate distinct amplification products.
• Integrated amplified products onto an electrochemical DNA chip with immobilized probes for automated hybridization, washing, and detection using the Genelyzer system.

Main Results:

• The novel system demonstrated high concordance with the existing nested PCR method.
• Positive and negative concordance rates were 100% (40/40) and 97.3% (117/120), respectively.
• The method proved to be simple, high-throughput, and significantly reduced the cost per sample.

Conclusions:

• The developed tag insertion primer and electrochemical DNA chip system offers a powerful tool for multisample detection.
• This technology is highly effective for diagnosing infectious agents in both animal and human samples.
• The system's efficiency, cost-effectiveness, and high-throughput capabilities make it a promising advancement in molecular diagnostics.