Method for selectively enriching and isolating microbial and optionally additional viral nucleic acids

摘要

The present invention relates to a method for selectively enriching and/or isolating microbial and optionally additionally viral nucleic acids from samples that contain a mixture of microbial cells, freely circulating nucleic acids and higher eukaryotic cells, and optionally additionally viruses, in a liquid, and to a kit for selectively enriching and/or isolating intracellular and extracellular microbial nucleic acids, and optionally additionally viral nucleic acids, from samples that contain a mixture of microbial and higher eukaryotic cells, freely circulating nucleic acids, in particular extracellular microbial nucleic acids, and optionally additionally viruses in a liquid.

主权项

1. A method for selectively accumulating or isolating microbial nucleic acids and viral nucleic acids from a sample that comprises a mixture of microbial cells, higher eukaryotic cells, free-circulating nucleic acids, and viruses in a liquid, comprising:
(1) substantially separating the liquid of the sample, including the free-circulating nucleic acids dissolved in the liquid and viruses, from the microbial cells and higher eukaryotic cells of the sample, (2) selectively lysing the eukaryotic cells from step (1), leaving the microbial cells intact, (3) substantially separating the eukaryotic lysate from step (2) from the intact microbial cells, (4) resuspending the intact microbial cells obtained in step (3), (5) lysing the microbial cells which are present in the suspension obtained in step (4), and (6) purifying or concentrating the free-circulating nucleic acids from the liquid obtained in step (1), viral nucleic acids from the viruses in the liquid obtained in step (1), and the intracellular microbial nucleic acids from the lysate obtained in step (5).