Rinderpest virus lineage differentiation using RT-PCR and SNAP-ELISA

Area of Science:

• Veterinary Virology
• Molecular Diagnostics
• Epidemiology

Background:

• Rinderpest virus (RPV) and Peste des petits ruminants virus (PPRV) are closely related morbilliviruses affecting ruminants.
• Accurate and rapid differentiation is crucial for disease control and outbreak management.
• Existing diagnostic methods can be time-consuming or require specialized equipment.

Purpose of the Study:

• To develop a novel diagnostic system for simultaneous detection and differentiation of RPV and PPRV.
• To enable identification of virus origin (wild-type vs. vaccine) and wild-type RPV outbreak source.
• To provide a faster, easier alternative to nucleotide sequencing or nested PCR for confirmation.

Main Methods:

• A combined Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Enzyme-Linked Immunosorbent Assay (ELISA) system was established.
• RT-PCR utilized a digoxigenin (Dig)-labeled primer.
• Hybridization employed virus-specific, biotin-labeled probes on a streptavidin-coated plate, with detection via anti-Dig and anti-mouse horseradish peroxidase conjugates.

Main Results:

• The system successfully detected and differentiated RPV from PPRV.
• Lineage-specific probes allowed identification of wild-type versus vaccine strains.
• Wild-type RPV samples could be traced to their likely outbreak origin.
• The assay provided semi-quantitative estimates of virus content.

Conclusions:

• The developed RT-PCR/ELISA assay offers a rapid, sensitive, and specific tool for RPV and PPRV diagnostics.
• This method simplifies confirmation of viral identity, replacing traditional sequencing or nested PCR.
• The assay's capabilities in strain differentiation and outbreak tracing enhance epidemiological surveillance and control strategies for morbillivirus infections in ruminants.