RNA from decades-old archival tissue blocks for retrospective studies.
Author(s): Mizuno T, Nagamura H, Iwamoto KS, Ito T, Fukuhara T, Tokunaga M, Tokuoka S, Mabuchi K, Seyama T
Publication: Diagn Mol Pathol, 1998, Vol. 7, Page 202-8
PubMed ID: 9917130 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to evaluate whether RNA could be isolated and amplified from autopsy tissues with varying postmortem intervals (PMI; up to 24 h) that had been formalin-fixed ( for 10 to 219 d) and stored as paraffin embedded tissue blocks for 10 to 40 years. Neoplastic liver and thyroid tissues were examined.
Summary of Findings:
The authors report successful amplification of c-BCR mRNA from 60% of liver biospecimens and 65% of thyroid biospecimens. Successful RT-PCR amplification of RNA extracted from archival formalin-fixed, paraffin-embedded specimens was dependent upon the length of storage, decreasing from a 80% success rate among specimens stored for 10 years to 40% after 40 years of storage. Similarly, RT-PCR analysis was adversely affected by increasing PMI, with RT-PCR success rate decreasing from 80% after a PMI to 0-2 h to 60% after 4-10 h. The duration of formalin fixation (10 to 219 d) did not significantly influence RT-PCR results. The authors report successful amplification of HCV in 39 of 74 liver carcinoma biospecimens and successful amplification of H4-RET in 2 of 19 thyroid carcinoma biospecimens.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
- Autopsy
Platform:
Analyte Technology Platform RNA RT-PCR DNA Southern blot Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 10-40 years
Preaquisition Postmortem interval 0-1 h 59 min
2 h- 3 h 59 min
4 h-9 h 59 min
10 h-23 h 59 min
>24 h
Biospecimen Preservation Time in fixative 10-20 d
21-30 d
31-60 d
61-219 d
Biospecimen Acquisition Biospecimen location Liver
Thyroid