A method for the detection of bacteria in from the blood samples derived with the aid of a realtime - pcr

摘要

A method for the detection of bacteria in derived from blood samples, comprisinga) of from blood samples derived,e) concentration of the derived from blood samples,f) extraction of the nucleic acids from the bacteria with the use of a lysis - buffer, an internal control - includes nucleic acid, and under the use of heated nuclease - free water for the elution,g) is obtained of the bacterial nucleic acid - extracts of step f),h) producing a pcr - master mixes without dna - polymerase, comprising a set of oligonucleotides of a respective sense - primer, an antisense - primer and a probe, wherein the one set of oligonucleotides is specific for bacterial nucleic acids, and further comprising a further probe, which is specific for the internal control - nucleic acid, and wherein the two probes are marked in each case with two fluorescent dyes,i) treatment of the pcr - master mixes of step h) and the obtained in step g) bacterial nucleic acid - extracts by addition of dnase i and subsequent addition of proteinase k,j) adding..