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J Histochem Cytochem. 2019 Mar;67(3):159-168. doi: 10.1369/0022155418819967. Epub 2018 Dec 18.

Cold Ischemia Score: An mRNA Assay for the Detection of Extended Cold Ischemia in Formalin-Fixed, Paraffin-Embedded Tissue.

Author information

1
Integrated Biobank of Luxembourg, Luxembourg.
2
National Cancer Institute, Bethesda, Maryland.
3
Van Andel Institute, Grand Rapids, Michigan.

Abstract

Although there are thousands of formalin-fixed paraffin-embedded (FFPE) tissue blocks potentially available for scientific research, many are of questionable quality, partly due to unknown preanalytical variables. We analyzed FFPE tissue biospecimens as part of the National Cancer Institute (NCI) Biospecimen Preanalytical Variables program to identify mRNA markers denoting cold ischemic time. The mRNA was extracted from colon, kidney, and ovary cancer FFPE blocks (40 patients, 10-12 hr fixation time) with 1, 2, 3, and 12 hr cold ischemic times, then analyzed using qRT-PCR for 23 genes selected following a literature search. No genes tested could determine short ischemic times (1-3 hr). However, a combination of three unstable genes normalized to a more stable gene could generate a "Cold Ischemia Score" that could distinguish 1 to 3 hr cold ischemia from 12 hr cold ischemia with 62% sensitivity and 84% specificity.

KEYWORDS:

; cancer; degradation; stability; transcript

PMID:
30562131
PMCID:
PMC6393842
DOI:
10.1369/0022155418819967
[Indexed for MEDLINE]
Free PMC Article

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