Standardized Recombinase Polymerase Amplification (RPA) Assay for Detecting Active Trypanosoma brucei evansi...

Area of Science:

• Veterinary Parasitology
• Molecular Diagnostics
• Zoonotic Diseases

Background:

• Surra, caused by Trypanosoma brucei evansi (T. b. evansi), significantly impacts livestock globally, leading to substantial economic losses.
• T. b. evansi, unlike T. brucei, spreads via mechanical vectors (biting flies), resulting in a wide and unpredictable distribution.
• Atypical human infections highlight T. b. evansi's potential as a zoonotic pathogen, underscoring the need for effective diagnostics.

Purpose of the Study:

• To address the need for rapid, accurate, and field-deployable diagnostic tests for Trypanosoma brucei evansi (T. b. evansi) infections.
• To evaluate recombinase polymerase amplification (RPA) as a suitable alternative to traditional PCR for T. b. evansi detection.
• To describe procedures for detecting active T. b. evansi infections based on application, resources, and sensitivity requirements.

Main Methods:

• Utilized recombinase polymerase amplification (RPA), an isothermal nucleic acid amplification technique.
• Explored various RPA result visualization methods: agarose gel electrophoresis (RPA-AGE), lateral flow assay (RPA-LFA), and real-time fluorescence (RPA-RT).
• Detailed specific procedures for detecting active T. b. evansi infections adaptable to different settings.

Main Results:

• RPA demonstrated potential as a simple, fast, and cost-effective method for pathogen detection.
• RPA is well-suited for use in resource-limited settings and field applications.
• The choice of RPA visualization method impacts sensitivity and applicability.

Conclusions:

• Recombinase polymerase amplification (RPA) provides a viable alternative to PCR for diagnosing Trypanosoma brucei evansi (T. b. evansi) infections.
• RPA's isothermal nature and adaptability make it ideal for field diagnostics in veterinary and potentially human health.
• Tailoring RPA procedures and visualization methods ensures effective detection based on specific diagnostic needs.