Whatman® FTA® Cards Performance for Ornithobacterium rhinotracheale DNA Amplification

Area of Science:

• Veterinary Microbiology
• Molecular Diagnostics
• Avian Pathology

Background:

• Ornithobacterium rhinotracheale (ORT) is an avian pathogen implicated in poultry respiratory disease.
• Effective sample handling is crucial for accurate diagnosis and disease surveillance.
• Traditional sample storage methods may pose risks for microorganism viability and transport safety.

Purpose of the Study:

• To evaluate the suitability of Whatman® Flinders Technology Associates (FTA®) cards for storing Ornithobacterium rhinotracheale (ORT) samples.
• To assess the efficacy of DNA amplification from ORT samples stored on FTA cards compared to sterile swabs.
• To determine the optimal conditions for hazard-free transport and storage of ORT samples for molecular analysis.

Main Methods:

• ORT culture dilutions were applied to FTA cards and sterile swabs.
• Samples were stored at room temperature, 4°C, and -20°C for up to 6 months.
• DNA was extracted at various time points, and real-time PCR targeting the 16S rRNA gene was performed.

Main Results:

• Real-time PCR amplification efficiency was comparable between FTA cards and swabs, showing dilution-dependent results.
• Threshold cycle values remained consistent across different storage temperatures and materials.
• Bacterial re-isolation was successful from swabs but not from FTA cards, indicating growth inhibition on FTA cards.

Conclusions:

• FTA cards provide an efficient and safe method for storing ORT samples for DNA amplification for at least 6 months.
• FTA cards offer the advantage of preventing microbial growth, ensuring hazard-free transport and storage.
• This method supports reliable molecular diagnostics for Ornithobacterium rhinotracheale in poultry.