Molecular techniques for the identification of bovine mastitis pathogens
Date
2012
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Abstract
Mastitis, one of the important production diseases of dairy cows, causes great economic losses to the dairy industry. Identification of mastitis pathogens using culturebased methods is often less accurate than genotypic methods. This is especially true in the case of coagulase-negative staphylococci (CNS), which were considered to be minor pathogens and now are gaining importance as the predominant pathogens in mastitis. In this work, a rapid assay, high-resolution melt analysis (HRMA) was tested for the species identification of bacterial mas ti tis pathogens including CNS targeting the variable regions of the 16S rDNA sequences. HRMA had an accuracy of 95% in species identification of common bacterial pathogens with only difficulty in identifying the streptococci due to heterogeneity observed in the PCR amplicons of Streptococcus uberis. HRMA could easily identify the most common CNS species, Staphylococcus chromogenes, which accounted for 68% of the CNS isolates in this study. Different genotypes of S. chromogenes also were identified using this technique. The remainder of the CNS were identified to species level by 16S rRNA, rpoB and/or tuf gene sequencing. This study demonstrates that real-time PCR and HRMA of 16S rDNA sequences can be employed successfully as an effective, low-cost technique for identification of common mastitis pathogens including S. chromogenes.
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Bibliography: p. 12-19, 45-49, 77-83, 95-100
Some pages are in colour.
Some pages are in colour.
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Citation
Ajitkumar, P. (2012). Molecular techniques for the identification of bovine mastitis pathogens (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/5022