Extraction, amplification and sequential hybridization of fungal cell DNA and process for detection of fungal cells in clinical material

A method for detection of fungal DNA in clinical material comprising:

• a) isolating predominantly intact fungal cells from whole blood by disintegrating blood cells by osmotic hemolysis and enzymatic digestion, and isolating intact fungal cells;
• b) isolating fungal DNA from the intact fungal cells by disintegrating the intact fungal cells by alkaline lysis and enzymatic treatment and isolating the fungal DNA;
• c) amplifying at least one segment of the fungal DNA to be detected by polymerase chain reaction;
• d) determining whether the nucleotide sequence of said at least one segment of fungal DNA is characteristic of a particular fungal strain and/or species, wherein said at least one segment of fungal DNA is hybridized with DNA probes specific for the particular fungal strain and/or species, said DNA probes being selected from the group consisting of SEQ ID-No:
• 3, SEQ ID-No:
• 4;
• SEQ ID-No:
• 5, SEQ ID-No-6, SEQ ID-No:
• 7 and SEQ ID-No:
• 8, and wherein a successful hybridization leads to identification of the fungal strain and/or species.

A method for detection of fungal DNA in clinical material comprising:

• a) isolating predominantly intact fungal cells from whole blood by disintegrating blood cells by osmotic hemolysis and enzymatic digestion, and isolating intact fungal cells;
• b) isolating fungal DNA from the intact fungal cells by disintegrating the intact fungal cells by alkaline lysis and enzymatic treatment and isolating the fungal DNA;
• c) amplifying at least one segment of the fungal DNA to be detected by polymerase chain reaction using at least one primer;
• d) determining whether the nucleotide sequence of said at least one segment of fungal DNA is characteristic of a particular fungal strain and/or species, wherein said at least one segment of fungal DNA is hybridized with DNA probes specific for the particular fungal strain and/or species, and wherein a successful hybridization leads to identification of the fungal strain and/or species.

A method for detection of fungal DNA in clinical material comprising:

• a) isolating predominantly intact fungal cells from whole blood by disintegrating blood cells and isolating intact fungal cells;
• b) isolating fungal DNA from the intact fungal cells by disintegrating the intact fungal cells and isolating the fungal DNA;
• c) amplifying at least one segment of the fungal DNA to be detected by polymerase chain reaction using at least one primer;
• and
• d) determining whether the nucleotide sequence of said at least one segment of fungal DNA is characteristic of a particular fungal strain and/or species, wherein said at least one segment of fungal DNA is hybridized with DNA probes specific for the particular fungal strain and/or species, and wherein a successful hybridization leads to identification of the fungal strain and/or species.