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Updated: Jun 3, 2026
Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications
Published on: June 29, 2020
Lyssavirus detection and typing using pyrosequencing.
Paola De Benedictis1, Cristian De Battisti, Laurent Dacheux
1OIE and National Collaborating Centre for Diseases at the Animal-Human Interface, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università 10, 35020 Legnaro, Padova, Italy. pdebenedictis@izsvenezie.it
A new pyrosequencing method accurately identifies rabies and related viruses. This cost-effective diagnostic approach enhances rabies detection in human and animal samples, improving public health surveillance.
Area of Science:
- Virology
- Molecular Diagnostics
- Public Health
Background:
- Rabies is a fatal zoonotic disease caused by rabies virus (RABV) and related lyssaviruses (RRVs).
- Increasing detection of diverse lyssaviruses in bat populations necessitates improved diagnostic tools.
- Current methods for lyssavirus identification can be costly and time-consuming.
Purpose of the Study:
- To develop and validate a rapid, cost-effective molecular assay for lyssavirus detection and differentiation.
- To apply pyrosequencing for the characterization of lyssaviruses targeting the nucleoprotein (N) gene.
- To establish a sensitive and specific diagnostic approach for rabies and related viruses.
Main Methods:
- A pyrosequencing protocol targeting the 3' terminus of the N gene was developed.
- A pan-lyssavirus one-step reverse transcription (RT)-PCR assay was integrated into the pyrosequencing workflow.
- Assay performance was validated using in vitro-transcribed RNA and serial dilutions of viruses.
- Results were confirmed using Sanger sequencing.
Main Results:
- The pyrosequencing assay successfully identified lyssavirus species in all tested samples.
- The developed RT-PCR assay demonstrated high analytical specificity (98.94%) and sensitivity (99.71%).
- Pyrosequencing offers a more affordable alternative for rapid lyssavirus typing compared to existing protocols.
Conclusions:
- Pyrosequencing provides a reliable and cost-effective method for rapid lyssavirus identification.
- The developed assay is suitable for detecting lyssaviruses in both human and animal samples.
- This approach can significantly enhance rabies surveillance and diagnosis globally.
