In situ hybridization detection of calcitonin mRNA in routinely fixed, paraffin-embedded tissue sections: a comparison of different types of probes combined with tyramide signal amplification.
Author(s): Qian X, Bauer RA, Xu HS, Lloyd RV
Publication: Appl Immunohistochem Mol Morphol, 2001, Vol. 9, Page 61-9
PubMed ID: 11277417 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to perform a comparative assessment of ISH sensitivity using different probes types (oligonucleotide, cDNA, cRNA) with and without post hybridization tyramide signal amplification in FFPE specimens.
Summary of Findings:
The authors report that cRNA probes are more sensitive than oligonucleotide or cDNA probes for detecting RNA molecules in FFPE tissue. As determined by both ISH and dot blot, tyramide signal amplification, in combination with cRNA probes, increased detection sensitivity 2- to 4-fold for calcitonin mRNA. While tyramide signal amplification also increased detection sensitivity when oligonucleotides or cDNA probes were used, nonspecific staining also increased.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform RNA In situ hybridization RNA Dot blot or slot blot Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) In situ hybridization Specific Type of probe cDNA probe
cRNA probe (riboprobe)
Oligonucleotide cocktail
Single oligonucleotide
In situ hybridization Specific Signal amplification Tyramide signal amplification
No signal amplification