Rapid and visual detection of Tacheng tick virus 1 using loop-mediated isothermal amplification technique

Area of Science:

• Virology
• Molecular Diagnostics
• Public Health

Background:

• Tacheng tick virus 1 (TcTV-1) is an emerging nairovirus transmitted by ticks, causing febrile illness in humans.
• Current diagnostic methods for TcTV-1 are limited, hindering effective surveillance and patient management.

Purpose of the Study:

• To develop and validate a rapid, sensitive, and specific loop-mediated isothermal amplification (LAMP) assay for TcTV-1 detection.

Main Methods:

• Primers were designed targeting the nucleocapsid protein (NP) gene of TcTV-1.
• Assay sensitivity and specificity were assessed using serial dilutions of plasmids and cross-reactivity testing with related viruses.
• The assay was validated on field-collected tick samples and compared with RT-qPCR.

Main Results:

• The TcTV-1 LAMP assay detected as few as 1x10^-1 copies/μL within 60 minutes at 65°C.
• The assay demonstrated high specificity, with no cross-reactivity to other tested nairoviruses.
• Validation on tick samples showed 100% sensitivity, specificity, PPV, NPV, and accuracy compared to RT-qPCR.

Conclusions:

• A sensitive, specific, and cost-effective TcTV-1 LAMP assay was successfully developed.
• This assay is a practical tool for field-based or resource-limited settings for TcTV-1 detection and surveillance.