A nested polymerase chain reaction assay to differentiate pestiviruses

Area of Science:

• Veterinary Virology
• Molecular Diagnostics
• Livestock Disease Control

Background:

• Pestiviruses cause substantial economic losses in the livestock sector.
• Current identification methods struggle to differentiate between bovine and ovine pestivirus isolates due to cross-reactivity.

Purpose of the Study:

• To develop a nested polymerase chain reaction (PCR) assay for precise identification and differentiation of three key pestivirus genotypes.
• To enable rapid characterization of viral isolates affecting cattle and sheep.

Main Methods:

• A nested RT-PCR assay was designed using consensus primers for initial amplification.
• Type-specific primers were employed in a second PCR round to generate unique DNA product sizes for each genotype.
• The assay was validated for distinguishing bovine viral diarrhea virus (BVDV) types I and II, and border disease virus (BDV).

Main Results:

• The developed nested PCR assay successfully amplified and differentiated between BVDV type I, BVDV type II, and BDV.
• The assay demonstrated the presence of all three pestivirus genotypes within ovine isolates.
• Rapid characterization of viral isolates was achieved.

Conclusions:

• The novel nested PCR assay provides a reliable and rapid method for distinguishing pestivirus genotypes in livestock.
• This diagnostic tool can improve disease surveillance and management strategies for BVDV and BDV.
• The findings confirm the circulation of multiple pestivirus genotypes in ovine populations.