DNA and RNA obtained from Bouin’s fixed tissues

• Published in: Journal of clinical pathology Vol. 58; no. 3; pp. 313 - 316
• Main Authors: Bonin, S, Petrera, F, Rosai, J, Stanta, G
• Format: Journal Article
• Language: English
• Published: London BMJ Publishing Group Ltd and Association of Clinical Pathologists 01-03-2005; BMJ; BMJ Publishing Group LTD; Copyright 2005 Journal of Clinical Pathology
• Subjects: Acetic Acid - pharmacology; Aqueous solutions; Biological and medical sciences; Biological Specimen Banks; Biopsy; Breast Neoplasms - genetics; Deoxyribonucleic acid; DNA; DNA Damage; DNA polymerase; DNA, Neoplasm - analysis; DNA, Neoplasm - drug effects; EGFR; epidermal growth factor receptor; Female; Fixatives - pharmacology; Formaldehyde - pharmacology; GAPDH; Gene expression; Genetic testing; human glyceraldehyde-3-phosphate dehydrogenase; Humans; Investigative techniques, diagnostic techniques (general aspects); Laboratories; Lymphoma; Medical sciences; Paraffin Embedding; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; PCR; Picrates - pharmacology; Polymerase chain reaction; Polymerase Chain Reaction - methods; Reverse Transcriptase Polymerase Chain Reaction - methods; reverse transcription; RNA, Neoplasm - analysis; RNA, Neoplasm - drug effects; Short Reports/Case Reports; Tissue Fixation - methods; transthyretin gene; TTR; Tissue; Anatomic pathology; RNA; DNA
• ISSN: 0021-9746; 1472-4146
• DOI: 10.1136/jcp.2004.016477

Background: The use in many countries of acid fixatives, such as Bouin’s solution, has limited the use of archival tissue for molecular analysis. An acidic environment is one of the main causes of DNA degradation. Moreover, RNA extraction is difficult in these types of fixed tissues. Aims: To amplify DNA and RNA from Bouin’s fixed tissues. Methods: DNA and RNA were extracted from 20 breast cancer samples that had been routinely fixed in Bouin’s fixative. Amplification of several genes using primers that produced amplicons of different lengths was carried out using the polymerase chain reaction (PCR) for DNA (with and without restoration) and reverse transcription PCR for RNA. Results: The acid environment of Bouin’s fixative damaged both DNA and RNA. However, amplification was successful when the amplicon length was reduced to about 80 bp for RNA and 100–200 bp for DNA, especially if submitted to DNA reconstruction procedures. Conclusions: It is possible to recover and analyse DNA and RNA from Bouin’s fixed and paraffin wax embedded tissues.