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Topic: Cell Free Plasma Dna As A Predictor Of Outcome In Severe Sepsis And Septic Shock

555 words - 2 pages

Introduction: The research is a clinical and correlation experiment performed by a Finland study group to find out the effect of increased concentration of cell free DNA found in 255 patients 18yrs or older who were seriously ill with whole body inflammatory state in both Intensive care unit and hospital mortality, as well as to see if there was a relation between cell free plasma DNA and organ failure.Procedure:Real time quantitative PCR assay for Beta globin gene was used to measure blood samples Age: 18yrs and older patients with severe sepsis or septic shock.Time: 72hrs after their blood was collected over a period of 4months in 24hrsInstrument: Centrifugation was used to separate the plasma fraction and the blood samples.Storage: stored at -20 degree C and later sent to Helsinki University Hospital, where they were stored at -80 degree C.Data Collection: Simplified acute physiologic score (SAPS II), acute physiology and chronic health evaluation (APACHE II), Sequential organ failure assessment (SOFA), lactate concentration and creatine clearance score using Cockcroft Gault formula were used to study the amount of cell free plasma DNA in survivors and non survivors in both ICU (intensive care unit) and Hospital patients.METHODS:1. The QIA amo DNA Blood Mini Kit (Qiagen) was used to extract DNA in order to ensure there weren’t any residual cells left behind.2. The DNA extracted produced the following sequence: forward primer 5’-GCA CCT….GAA-3’. Reverse primer 5’CAC CAA……TCA-3’ as well as a single labeled fluorescent MGB probe 5’-FAM-TCT….MGB-NFQ. (Minor grove binding molecule and Nonfluorescent quencher molecule respectively).Precision of Data:The data was ran 8times in multiple duplicates to ensure precision of the real time quantitative PCR...

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