Quantitative PCR high-resolution melting (qPCR-hRM) curve analysis, a new approach to rapid detection and differentiation of bovine papillomavirus detected in equine sarcoids.

This study introduces and evaluates a new diagnostic method that employs quantitative PCR and High-Resolution Melting (qPCR-HRM) curve analysis for quick differentiation of bovine papillomavirus patterns known as BPV-1 or BPV-2, found in horse sarcoids. Additionally, the research includes a phylogenetic examination of the complete coding sequence of the E5 gene of BPV in these equine skin lesions.

Methodology and Results

• The researchers acquired samples from 38 skin lesions from 27 horses, all of which were clinically diagnosed as sarcoids, a type of skin growth.
• These samples were subject to molecular examinations where they were tested for the presence of BPV DNA.
• All the samples were positive for BPV DNA; subsequently, qPCR-HRM analysis helped identify 6 specimens (16% of samples) as BPV-1 “wild”, 24 (63% of samples) as BPV-1 “European”, and 8 (21% of samples) as a “variant” of BPV E5 ORF partial coding sequence.

Phylogenetic Analysis

• The research also involved a phylogenetic analysis which is essentially a study of evolutionary relationships based on nucleotide sequences of E2 ORF partial coding sequence and E5 ORF complete coding sequence.
• This analysis was conducted on 7 specimens whose sequences were published in GenBank and identified as different variants of BPV-1 and BPV-2.

Novel Findings

• Upon analyzing the amino acid sequences of E5 ORF complete CDS, the researchers identified a variant in the isolate 10PL.
• The variation found in the isolate 10PL is a new variant of the E5 ORF complete coding sequence of BPV-1 found in equine sarcoid in Poland, making it a significant discovery in this research.

Overall, this paper describes the significance and effectiveness of a new diagnostic model combining qPCR and HRM for BPV differentiation in equine sarcoids. Further, it highlights the discovery of a previously unknown variant of BPV-1 in Polish equine sarcoids.