Comparison of a Selection of Rapid Automated DNA and RNA Extraction Technologies for Detection of Somatic or Constitutional Gene Abnormalities in Cancer Diagnosis

Abstract

Molecular analyses of large numbers of patient samples are increasingly used for diagnostic applications as well as for understanding cancer biology. Their accuracy depends on the quality and quantity of nucleic acids extracted from human cells or tissues. To optimize these preanalytical steps, we evaluated several automated technologies for nucleic acid purification from clinical samples. Three automated platforms were compared. DNA was extracted from peripheral blood leukocytes from five normal individuals, and its quality was assessed by D-HPLC and sequencing after PCR. Clinical samples from acute leukemia patients were used for automated RNA extractions; results were compared to our standard manual technique. RNA qualification was done using capillary gel electrophoresis and analysis of the Abelson gene transcript by real-time PCR. One robot produced higher total output for both DNA and RNA. While the quality of DNAs obtained from the three workstations allowed implementation of their analysis for detection of germinal mutations, important differences were observed in the quality of RNAs. One robot isolated RNA with similar quality and quantity to the manual technique, but the resulting products displayed low concentrations. A robust technique had therefore to be evaluated and validated to allow the implementation of this workstation within the daily diagnostic practices. This study is of interest at a time when hospital-based laboratories are bringing molecular signatures to the clinic. 2

Total RNA extraction method and Prunus species influence the detection of Plum pox potyvirus by real-time RT-PCR

“…The use of KingFisher instrument is automated and fast, it saves time and money. The financial disadvantage is the initial investment into the instrument but it can be used for many different applications (for DNA, RNA and protein extraction) (Lassailly et al, 2007;Zhao et al, 2008;Josefsen et al, 2007).…”

RNA Extracted From Blood Samples With a Rapid Automated Procedure Is Fit for Molecular Diagnosis or Minimal Residual Disease Monitoring in Patients With a Variety of Malignant Blood Disorders

“…This device was previously validated for its ability to improve RNA concentration without deteriorating the quality of subsequent RQ-PCR analyses. 1 RNAs were eluted in a fi nal volume of 15 μL of RNase-free water.…”