Molecular Diagnosis of Phlebovirus riftense Infection Using an L-Segment-Based Real-Time RT-PCR Method

Area of Science:

• Veterinary Medicine
• Molecular Biology
• Infectious Diseases

Background:

• Rift Valley fever (RVF) is a major vector-borne zoonotic disease impacting ruminants and humans in Africa and the Arabian Peninsula.
• Accurate diagnosis of RVF infection is crucial, especially during outbreaks, relying on timely sample transport and appropriate laboratory testing.
• Existing diagnostic methods include cell culture and molecular techniques, with real-time RT-PCR assays being the most rapid and sensitive.

Purpose of the Study:

• To describe a one-step real-time duplex RT-PCR technique for Rift Valley fever virus (RVFV) detection and quantification.
• To provide a detailed method for rapid and sensitive molecular diagnosis of RVF during outbreaks.

Main Methods:

• The study details a one-step real-time duplex RT-PCR assay targeting the L segment of the RVFV.
• This method is designed for rapid detection and quantification of viral particles.
• Emphasis is placed on sample quality, laboratory capacity, and the period of viremia for optimal results.

Main Results:

• Conventional and duplex/multiplex real-time RT-PCR assays are identified as the most rapid and sensitive molecular tests for RVFV detection.
• The described one-step real-time duplex RT-PCR offers a sensitive approach for RVFV diagnosis.
• Accurate interpretation requires validation with positive and negative controls.

Conclusions:

• The one-step real-time duplex RT-PCR is a valuable tool for the rapid and sensitive diagnosis of Rift Valley fever virus (RVFV).
• Effective implementation requires careful sample handling and interpretation of results in conjunction with controls.
• This technique aids in managing RVF outbreaks in affected regions.