Homogenous M13 bacteriophage quantification assay using switchable lanthanide fluorescence probes
| Autor: | Ari Lehmusvuori, Julius Manninen, Tuomas Huovinen, Tero Soukka, Urpo Lamminmäki |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 53, Iss 5, Pp 301-303 (2012) |
| Druh dokumentu: | article |
| ISSN: | 00001139 1940-9818 0736-6205 |
| DOI: | 10.2144/0000113954 |
| Popis: | We have developed a rapid and reliable bacteriophage quantification method based on measurement of phage single-stranded DNA (ssDNA) using switchable lanthanide chelate complementation probes. One oligonucleotide probe contains a non-fluorescent lanthanide ion carrier chelate and another probe is labeled with a light absorbing antenna ligand. Hybridization of the non-fluorescent complementation probes in adjacent positions on the released bacteriophage ssDNA leads to high local concentrations of the lanthanide ion carrier chelate and the antenna ligand, inducing formation of a fluorescent lanthanide chelate complex. This method enables monitoring of bacteriophage titers in a 20 min assay with a dynamic range of 109–1012 cfu/mL in a microtiter well format. While designed for titering filamentous bacteriophage used in phage display, our method also could be implemented in virological research as a tool to analyze ssDNA virus reproduction. |
| Databáze: | Directory of Open Access Journals |
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