Loop-Mediated Isothermal Amplification on Paper Microfluidic Chips for Highly Sensitive and Specific Zika Virus...

Area of Science:

• Virology
• Molecular Biology
• Diagnostics

Background:

• Zika virus (ZIKV) poses significant risks, particularly for pregnant women, necessitating rapid diagnostic tools.
• Current detection methods may lack portability and ease of use for field applications.
• Point-of-care testing is crucial for controlling ZIKV transmission in affected regions.

Purpose of the Study:

• To develop a simple, portable, and user-friendly method for ZIKV RNA detection.
• To enable rapid, on-site diagnosis of Zika virus infections.
• To provide a tool for preventing the spread of ZIKV.

Main Methods:

• A reverse transcription isothermal loop-mediated amplification (RT-LAMP) assay was developed.
• Phenol red was used as a colorimetric indicator for successful amplification.
• Amplification was detected visually using smartphone cameras under ambient light.

Main Results:

• The RT-LAMP method detected ZIKV RNA in blood, urine, and tap water samples.
• Detection of a single viral RNA molecule per μL was achieved in as little as 15 minutes.
• The assay demonstrated 100% sensitivity and 100% specificity in blood and tap water.
• 100% sensitivity and 67% specificity were observed in urine samples.

Conclusions:

• This RT-LAMP platform offers a rapid and sensitive method for ZIKV detection.
• The smartphone-based colorimetric readout facilitates point-of-care and field-based diagnostics.
• The technology shows potential for detecting other viruses like SARS-CoV-2, enhancing diagnostic capabilities.