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Safe and cost-effective protocol for shipment of samples from Foot-and-Mouth Disease suspected cases for laboratory
1Laboratoire de Santé Animale de Maisons-Alfort, Laboratoire OIE de référence Fièvre Aphteuse, UMR Virologie 1161, Université Paris-Est, Anses, Maisons-Alfort, France.
Abstract:
An essential step towards the global control and eradication of foot-and-mouth disease (FMD) is the identification of circulating virus strains in endemic regions to implement adequate outbreak control measures. However, due to the high biological risk and the requirement for biological samples to be shipped frozen, the cost of shipping samples becomes one of major obstacles hindering submission of suspected samples to reference laboratories for virus identification. In this study, we report the development of a cost-effective and safe method for shipment of FMD samples. The protocol is based on the inactivation of FMD virus (FMDV) on lateral flow device (LFD, penside test routinely used in the field for rapid immunodetection of FMDV), allowing its subsequent detection and typing by RT-PCR and recovery of live virus upon RNA transfection into permissive cells. After live FMDV collection onto LFD strip and soaking in 0.2% citric acid solution, the virus is totally inactivated. Viral RNA is still detectable by real-time RT-PCR following inactivation, and the virus strain can be characterized by sequencing of the VP1 coding region. In addition, live virus can be rescued by transfecting RNA extract from treated LFD into cells. This protocol should help promoting submission of FMD suspected samples to reference laboratories (by reducing the cost of sample shipping) and thus characterization of FMDV strains circulating in endemic regions.
Insights
A new method inactivates foot-and-mouth disease virus (FMDV) on lateral flow devices (LFDs) for safe, cost-effective sample shipping. This allows for virus detection, typing, and even recovery, aiding global FMD control efforts.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Epidemiology
Background:
- Global control of foot-and-mouth disease (FMD) requires identifying circulating virus strains.
- Shipping frozen biological samples for FMD virus identification is costly and poses a high biological risk, hindering sample submission.
Purpose of the Study:
- To develop a cost-effective and safe method for shipping FMD samples.
- To enable virus identification and characterization from suspected FMD outbreaks.
Main Methods:
- Inactivation of FMD virus (FMDV) on lateral flow devices (LFDs) using citric acid.
- Detection and typing of viral RNA using real-time RT-PCR and VP1 gene sequencing.
- Recovery of infectious live virus via RNA transfection into permissive cells.
Main Results:
- FMDV was effectively inactivated on LFDs after treatment with 0.2% citric acid.
- Viral RNA remained detectable by real-time RT-PCR post-inactivation.
- Virus strain characterization through VP1 sequencing was successful.
- Live virus was rescued from treated LFD samples.
Conclusions:
- The developed protocol offers a safe and economical alternative for FMD sample shipment.
- This method facilitates the submission of suspected FMD samples to reference laboratories.
- Improved sample submission will enhance the characterization of FMDV strains in endemic regions, aiding disease control.
