Simultaneous DNA/RNA Isolation from Plasma or Serum
Comparison chemagic Kits vs. Column Technique
The chemagic Viral DNA/RNA Kit is designed for simultaneous purification of viral DNA and RNA from 200 µl serum or plasma samples. To increase the sensitivity of detection the sample size is scalable. The following gel image shows a comparison of M-PVA Magnetic Bead based technology to conventional column methods.
Fig.: HBV Detection. PCR products amplified from DNA isolated either with the chemagic Viral DNA/RNA Kit (upper row) or a column based kit (lower row).
(1) - (22) PCR products generated from isolated DNA using sera from different patients.
(M): DNA marker. Bands of the upper row and the lower row with the same number are amplificates using eluates obtained from the serum of the same patient.
(N) Negative control.
chemagic Viral DNA/RNA Kits on chemagic Automation
Applied on chemagic Automation, especially regarding large volumes, the kits provide outstanding advantages. The protocol allows direct handling of plasma or serum samples without any intermediary centrifugation or alternative concentration steps. DNA- and RNA-viruses are prepared simultaneously in one extraction.
Sensitivity Data
DNA/RNA was isolated simultaneously from 9.6 ml minipool plasma samples (96 donors, pool of 100 µl aliquots). The validations were performed at renowned German Red Cross institutes. Different inhouse and commercial TaqMan or Lightcycler assays were used for the detection of viral NAs. A probit analysis of the results revealed the minimal virus concentrations detectable with 95 % probability (see table).
HAV1 | HBV2 | HCV2 | HIV2 | Parvo B193 | |
Detection limits | 2.6 | 0.7 | 5.0 | 33 | 4.6 |
Data kindly provided by
- DRK-Blutspendedienst West gGmbH, Zentrallabor Hagen;
- DRK-Blutspendedienst Sachsen gGmbH, Institut für Transfusionsmedizin Plauen;
- DRK-Blutspendedienst Hessen gGmbH, Institut für Transfusionsmedizin Frankfurt.
