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A method for detecting variability arising from errors in sample processing of paraffin-embedded tissue for DNA

J B Hendricks1, N S Hardt, E J Wilkinson

  • 1Department of Pathology, University of Florida College of Medicine, Gainesville.

Archives of Pathology & Laboratory Medicine
|November 1, 1993
PubMed
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A new method uses tonsil tissue controls to manage DNA content analysis variability in paraffin-embedded tissues. This approach reduces errors and improves the accuracy of clinical specimen results.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Histology

Background:

  • Variability in DNA content analysis of paraffin-embedded tissues can stem from inconsistent sample preparation.
  • Standard protocols for embedding and sectioning tissue may introduce errors affecting analytical results.

Purpose of the Study:

  • To develop and validate a method for controlling sample preparation variability in DNA content analysis of paraffin-embedded tissues.
  • To establish a reliable control system to ensure the accuracy and reproducibility of DNA content analysis.

Main Methods:

  • Human tonsil tissue sections were used as daily controls for DNA content analysis over 20 days.
  • A rejection rule (12s) was implemented: if the tonsil G0/G1 peak coefficient of variation (CV) exceeded 2 standard deviations, samples were reprocessed.

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  • Instrument performance and staining quality were verified using external controls.

    • Main Results:

    • The implemented rejection rule resulted in approximately 6% of sample runs requiring reprocessing.
    • Reprocessing runs with the tonsil control within acceptable ranges improved the G0/G1 peak CV of clinical specimens by 25% on average.
    • The method effectively detected decreased peak resolution due to sample processing errors.

    Conclusions:

    • A tonsil tissue control method effectively manages variability in DNA content analysis of paraffin-embedded tissues.
    • This control strategy enhances the reliability and accuracy of results from clinical specimens.
    • The method provides a valuable tool for detecting and correcting errors in sample preparation, crucial for diagnostic accuracy.