Related Experiment Video
Updated: May 18, 2026
Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR (RS-PCR)
Published on: November 4, 2016
Technical note: Identification of Prototheca species from bovine milk samples by PCR-single strand conformation
P Cremonesi1, F Pozzi, M Ricchi
1Istituto di Biologia e Biotecnologia Agraria, Consiglio Nazionale delle Ricerche (CNR), Lodi, Italy. cremonesi@ibba.cnr.it
Abstract:
We report the development of a PCR-single strand conformation polymorphism (SSCP) method to identify Prototheca spp. responsible for bovine mastitis: P. zopfii and P. blaschkeae. The method was set up using reference strains belonging to P. zopfii genotype 1, P. zopfii genotype 2, and P. blaschkeae as target species and P. stagnora, and P. ulmea as negative controls. The assay was applied on 50 isolates of Prototheca spp. isolated from bovine mastitic milk or bulk-tank milk samples, and all isolates were identified as P. zopfii genotype 2. We conclude that the described PCR-SSCP approach is accurate, inexpensive, and highly suitable for the identification of P. zopfii genotype 2 on field isolates but also directly on milk, if preceded by a specific DNA extraction method.
Insights
A new PCR-single strand conformation polymorphism (SSCP) method accurately identifies Prototheca species causing bovine mastitis. The assay successfully identified all 50 field isolates as Prototheca zopfii genotype 2.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
- Dairy Science
Background:
- Prototheca species are emerging pathogens causing bovine mastitis.
- Accurate identification of Prototheca species is crucial for effective disease management.
- Current diagnostic methods may lack specificity or efficiency for field applications.
Purpose of the Study:
- To develop and validate a Polymerase Chain Reaction-Single Strand Conformation Polymorphism (PCR-SSCP) assay.
- To differentiate between Prototheca zopfii (genotypes 1 and 2) and Prototheca blaschkeae.
- To assess the utility of the PCR-SSCP method for identifying Prototheca species in bovine mastitis cases.
Main Methods:
- Development of a PCR-SSCP assay using reference strains of Prototheca zopfii (genotypes 1 and 2) and P. blaschkeae.
- Inclusion of P. stagnora and P. ulmea as negative controls.
- Application of the assay to 50 Prototheca isolates from bovine mastitic milk and bulk-tank milk samples.
Main Results:
- The PCR-SSCP method was successfully established using reference strains.
- All 50 field isolates of Prototheca spp. were identified as P. zopfii genotype 2.
- The assay demonstrated high accuracy in identifying the target species.
Conclusions:
- The developed PCR-SSCP approach is accurate, cost-effective, and suitable for identifying P. zopfii genotype 2.
- The method is applicable to field isolates and potentially directly to milk samples with appropriate DNA extraction.
- This diagnostic tool can aid in understanding the epidemiology of Prototheca-associated bovine mastitis.
