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Detection of hepatitis C virus RNA in more than 20-year-old paraffin-embedded liver tissue.

Author(s): Soguero C, Ribalta T, Campo E, Sánchez-Tapies JM, Sáiz JC, Bruguera M

Publication: Lab Invest, 1999, Vol. 79, Page 365-6

PubMed ID: 10092073 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to optimize RNA extraction for reverse transcription polymerase chain reaction (RT-PCR) detection of hepatitis C viral RNA in formalin-fixed, paraffin-embedded (FFPE) liver specimens stored for varying durations (1 to 27 years).

Conclusion of Paper

The authors report that extended proteinase K digestion for 4 days at 42 degrees C enabled detection of hepatitis C viral RNA in 100% of FFPE liver specimens that had been stored for up to 10 years old and 70% of liver specimens that had been stored between 13 and 27 years.

Studies

  1. Study Purpose

    The purpose of this study was to evaluate two different proteinase K digestion protocols for extraction and identification of hepatitis C RNA in FFPE specimens stored for less than 10 years.

    Summary of Findings:

    Extended proteinase K incubation (42 degrees C for 4 days) increased the successful detection of the cDNA target via RT-PCR for both amplicon sizes (171 bp, 286 bp) compared to a shorter incubation at 62 degrees C for 5 hours.

    Biospecimens
    Preservative Types
    • Formalin
    Diagnoses:
    • Hepatitis
    Platform:
    AnalyteTechnology Platform
    RNA RT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Analyte Extraction and Purification Protein digestion Proteinase K (5 h, 62 degrees C)
    Proteinase K (4 d, 42 degrees C)
    RT-PCR Specific Length of gene fragment 171 bp
    286 bp
    View more
  2. Study Purpose

    The purpose of this study was to evaluate the ability to detect hepatitis C RNA via RT-PCR in formalin-fixed paraffin blocks that had been stored for extended periods of time (up to 27 years).

    Summary of Findings:

    When RNA was extracted according to the extended proteinase K digestion, hepatitis C RNA was identified in 100% of specimens that had been stored for 1 to 5 years for both cDNA amplicon sizes. For specimens stored for 6 to 10 years, RT-PCR detection of the 171 bp cDNA fragment was successful for 100% of specimens, while detection of the larger 286 bp fragments was successful for 7 of 8 specimens analyzed. Identification of the 171 bp cDNA fragment of hepatitis C RNA was successful in 70% of specimens stored for 13 to 27 years.

    Biospecimens
    Preservative Types
    • Formalin
    Diagnoses:
    • Hepatitis
    Platform:
    AnalyteTechnology Platform
    RNA RT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage duration 1-5 yr
    6-10 yr
    13-27 yr
    RT-PCR Specific Length of gene fragment 171 bp
    286 bp
    View more

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