| Autor: |
Ricchi, M., Cammi, G., Garbarino, C.A., Buzzini, P., Belletti, G.L., Arrigoni, N. |
| Předmět: |
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| Zdroj: |
Journal of Applied Microbiology; Jan2011, Vol. 110 Issue 1, p27-34, 8p, 3 Charts, 2 Graphs |
| Abstrakt: |
The study describes the development of a simple and rapid tool to identify yeast-like microalgae belonging to the genus Prototheca. The method, based on two-step Real Time PCR reaction followed by DNA Resolution Melting Analysis (qPCR/RMA), has been developed using reference strains belonging to both pathogenic ( P. zopfii genotype 2, P. wickerhamii and P. blaschkeae) and non -pathogenic species ( P. zopfii genotype 1, P. stagnora and P. ulmea). In order to validate the method, seventy recently isolated Prototheca strains were thus tested in parallel with both the first qPCR/RMA and the conventional genotype-specific PCR assay: they were classified as P. zopfii genotype 1, P. zopfii genotype 2 and P. blaschkeae, with a perfect accordance between the two above methodologies. Furthermore, we used the second qPCR/RMA to identify the other species ( P. stagnora, P. ulmea and P. wickerhamii), which cannot be discriminated by conventional PCR assay. The assay two-step Real Time PCR is accurate, robust, cost-effective and faster than auxonographical, biochemical or conventional molecular biology methods. the rapid and high throughout two-step qPCR/RMA tool can be usefully used for the identification of clinical and environmental Prototheca species into the framework of the diagnosis of animal (e.g. bovine mastitis) or human protothecosis. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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