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Related Experiment Videos

Specimen stability for DNA-based diagnostic testing

D H Farkas1, A M Drevon, F L Kiechle

  • 1Department of Clinical Pathology, William Beaumont Hospital, Royal Oak, Michigan 48073-6769, USA.

Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B
|December 1, 1996
PubMed
Summary
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DNA stability in clinical samples is crucial for molecular diagnostics. Freezing or refrigerating leukocytes and freezing solid tissue are preferred storage methods, with isolated DNA remaining stable for years.

Area of Science:

  • Molecular diagnostics
  • Clinical pathology
  • Biochemistry

Background:

  • Increasing use of molecular diagnostic testing in clinical settings.
  • Need for data on DNA stability in clinical specimens for accurate test performance and interpretation.

Purpose of the Study:

  • To determine DNA stability in peripheral blood and solid tissue under various storage conditions.
  • To evaluate DNA stability during storage at 4°C for up to 3 years.
  • To assess the impact of DNA degradation on Southern hybridization and polymerase chain reaction (PCR) analyses.

Main Methods:

  • Assayed DNA quality and yield using spectrophotometric absorbance and gel electrophoresis.
  • Assessed DNA suitability for Southern hybridization and PCR.
  • Stored peripheral blood leukocytes and solid tissue under different conditions (4°C, freezing) for up to 3 years.

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Main Results:

  • Freezing or refrigeration of separated leukocytes is optimal for short- to intermediate-term storage.
  • Freezing is the preferred method for solid tissue storage.
  • Isolated DNA demonstrated stability at 4°C for at least 3 years.
  • DNA degradation occurred inconsistently in specimens, but PCR and Southern hybridization were often still possible.

Conclusions:

  • Freezing or refrigeration protocols enhance DNA stability in clinical specimens.
  • Degraded DNA samples may still yield interpretable results via PCR or Southern hybridization.
  • Results suggest a more flexible approach to specimen requirements in molecular pathology.