nucleic acid molecule to methods for detecting fungi in clinical material and for determining the sequence of ribosomal fungal genes and to kits for carrying out these methods application_number US 42615903 A description a blood sample is taken from patients to be investigated for a fungal infection the erythrocytes from the whole blood are lysed hypotonically with rclb buffer 10 mm tris ph 7 6 5 mm mgcl 2 10 mm nacl this is followed by enzymatic lysis of the leukocytes with wclb buffer 10 mm tris ph 7 6 10 mm edta 50 mm nacl 0 2 sds 200 u03bcg of proteinase k per ml at 65 u00b0 c for 45 minutes this ensures detection also of fungal dna derived from fungal cells located inside blood cells the samples are pelleted and incubated with 50 mm naoh at 95 u00b0 c for 10 minutes this is followed by neutralization with 1 m tris ph 7 0 followed by treatment with recombinant lyticase sigma deissenhofen germany in a buffer which contains 1 u of lyticase per 100 u03bcl 50 mm tris ph 7 5 1 mm edta and 0 2 u03b2 mercaptoethanol at 37 u00b0 c for 45 minutes in order to form spheroplasts after centrifugation at 5000 g the supernatant which contains the human dna and proteins is decanted off and the pellets are treated with 1 m tris edta and 10 sds at 65 u00b0 c for 30 minutes to lyse the spheroplasts 5 m potassium acetate is then added and the samples are incubated at u2212 20 u00b0 c for 30 minutes to precipitate proteins after a further centrifugation step at 100 g for 20 minutes the dna is precipitated from the supernatant with cold isopropanol the dna is purified with 70 strength.