A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Homogenous M13 bacteriophage quantification assay using switchable lanthanide fluorescence probes
Tekijät: Lehmusvuori A, Manninen J, Huovinen T, Soukka T, Lamminmäki U
Kustantaja: BIOTECHNIQUES OFFICE
Julkaisuvuosi: 2012
Journal: Biotechniques
Tietokannassa oleva lehden nimi: BIOTECHNIQUES
Lehden akronyymi: BIOTECHNIQUES
Numero sarjassa: 5
Vuosikerta: 53
Numero: 5
Aloitussivu: 301
Lopetussivu: 303
Sivujen määrä: 3
ISSN: 0736-6205
DOI: https://doi.org/10.2144/0000113954
Tiivistelmä
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 10(9)-10(12) 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.
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 10(9)-10(12) 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.
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