|WANG, DEGUO - Xuchang University|
Submitted to: International Journal of Environmental Research and Public Health
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/8/2015
Publication Date: 5/26/2015
Publication URL: http://handle.nal.usda.gov/10113/61891
Citation: Wang, D., Liu, Y. 2015. Development of primer sets for loop-mediated isothermal amplification that enables rapid and specific detection of Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae. International Journal of Environmental Research and Public Health. 12(6):5735-5742.
Interpretive Summary: Bovine mastitis (BM) is a persistent and inflammatory reaction of the udder tissue in dairy cattle. Bovine mastitis is caused by bacteria known as Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae. Since there is no visible sign of infection when cow is infected with subclinical mastitis, a diagnostic method is needed for the control of infection in the herd. We have developed a Loop mediated isothermal amplification (LAMP) assay to detect S. dysgalactiae, S. uberis, and S. agalactiae. LAMP assay is a nucleic acid amplification technique in which the target sequence is amplified at a constant temperature using two or three sets of primers and a polymerase. Our results show that the LAMP assay has the potential to be used as a simple screening assay in the field or at the clinical setting.
Technical Abstract: Streptococcus dysgalactiae, Streptococcus uberis and Streptococcus agalactiae are the three main pathogens causing bovine mastitis, with great losses to the dairy industry. Rapid and specific loop-mediated isothermal amplification methods (LAMP) for identification and differentiation of these three pathogens are not available. With the 16S rRNA gene and 16S-23S rRNA intergenic spacers as targets, four sets of LAMP primers were designed for identification and differentiation of S. dysgalactiae, S. uberis and S. agalactiae. The detection limit of all four LAMP primer sets were 0.1 pg DNA template per reaction, the LAMP method with 16S rRNA gene and 16S-23S rRNA intergenic spacers as the targets can differentiate the three pathogens, which is potentially useful in epidemiological studies.