Homogenous M13 bacteriophage quantification assay using switchable lanthanide fluorescence probes | Zendy

Ari Lehmusvuori | Zendy; Julius Manninen | Zendy; Tuomas Huovinen | Zendy; Tero Soukka | Zendy; Urpo Lamminmäki | Zendy

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Homogenous M13 bacteriophage quantification assay using switchable lanthanide fluorescence probes

Author(s) -

Ari Lehmusvuori,

Julius Manninen,

Tuomas Huovinen,

Tero Soukka,

Urpo Lamminmäki

Publication year - 2012

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/0000113954

Subject(s) - bacteriophage , lanthanide , fluorescence , oligonucleotide , chelation , ligand (biochemistry) , myoviridae , chemistry , molecular probe , complementation , dna , biophysics , biology , ion , biochemistry , escherichia coli , inorganic chemistry , physics , receptor , organic chemistry , gene , quantum mechanics , phenotype

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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