Detection of mycoplasmas in goat milk by flow cytometry

Autor: Hazel M. Davey, P. Assunção, José B. Poveda, Nuno T. Antunes, Carlos M. Ruiz de Galarreta, Christian de la Fe, Ana S. Ramírez, Rubén S. Rosales
Rok vydání: 2007
Předmět:
Zdroj: Cytometry Part A. :1034-1038
ISSN: 1552-4930
1552-4922
DOI: 10.1002/cyto.a.20476
Popis: The detection of mycoplasma in milk can be performed by either culture techniques or polymerase chain reaction (PCR) based methods. Although PCR can reduce the average diagnostic time to 5 h in comparison with the several days for the isolation of the agent, there is still a need to develop methods, which could give earlier results. For this purpose, we tested the ability of flow cytometry (FC) to detect mycoplasmas in milk samples. Milk samples inoculated with four different mycoplasmas, Mycoplasma agalactiae, Mycoplasma putrefaciens, Mycoplasma capricolum subsp. Capricolum ,o rMycoplasma mycoides subsp. mycoides large-colony type, known to cause contagious agalactia in goats, were stained with the DNA stain SYBR Green I and analyzed by FC. Three goat milk samples, from which mycoplasmas have been isolated in broth medium were also analyzed. All mycoplasmas were easily distinguished from debris of milk samples, but it was not possible to distinguish between the different mycoplasma species. In our conditions, the detection limit of the technique was of the order of 10 3 –10 4 cells ml 21 . Furthermore, mycoplasmas were also distinguished from Staphylococcus aureus. FC together with SYBR Green I was able to distinguish between mycoplasma cells and debris present in milk samples and gave results in 20–30 min. This is an important first step in developing a robust, routine flow cytometric method for the detection of mycoplasmas in milk samples. ' 2007 International Society for Analytical Cytology
Databáze: OpenAIRE
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