Lessons from the Field

Practical tips to using formalin-fixed paraffin-embedded tissue archives for molecular diagnostics in a South African setting

Barbara S. van Deventer, Lorraine du Toit-Prinsloo, Chantal van Niekerk
African Journal of Laboratory Medicine | Vol 11, No 1 | a1587 | DOI: https://doi.org/10.4102/ajlm.v11i1.1587 | © 2022 Barbara S. van Deventer, Lorraine du Toit-Prinsloo, Chantal van Niekerk | This work is licensed under CC Attribution 4.0
Submitted: 16 March 2021 | Published: 23 June 2022

About the author(s)

Barbara S. van Deventer, Department of Forensic Medicine, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa
Lorraine du Toit-Prinsloo, Department of Forensic Medicine, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa
Chantal van Niekerk, Department of Chemical Pathology, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa


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Abstract

Background: Formalin-fixed paraffin-embedded (FFPE) tissue archives in hospitals, biobanks, and others offer a vast collection of extensive, readily available specimens for molecular testing. Unfortunately, the use of tissue samples for molecular diagnostic applications is challenging; thus, the forensic pathology FFPE tissue archives in Africa have been a largely unexploited genetic resource, with the usability of DNA obtainable from these samples being unknown.

Intervention: The study, conducted from January 2015 to August 2016, determined the usefulness of FFPE tissue as a reliable source of genetic material for successful post-mortem molecular applications and diagnostics. Formalin-fixed paraffin-embedded tissue samples were collected and archived from autopsies conducted over 13 years in the forensic medicine department of the University of Pretoria (Pretoria, South Africa). Deoxyribonucleic acid from FFPE tissue samples and control blood samples was amplified by high-resolution melt real-time polymerase chain reaction before sequencing. The procurement parameters and fixation times were compared with the quantity and quality of the extracted DNA and the efficiency of its subsequent molecular applications.

Lessons learnt: This study has shown that FFPE samples are still usable in molecular forensics, despite inadequate sample preparation, and offer immense value to forensic molecular diagnostics.

Recommendations: FFPE samples fixed in formalin for more than 24 h should still be used in molecular diagnostics or research, as long as the primer design targets amplicons not exceeding 300 base pairs.

 


Keywords

Autopsy; deoxyribonucleic acid; formalin-fixed paraffin-embedded tissue; formalin-fixed paraffin-embedded tissue archive; high-resolution melt analysis; molecular diagnostics; polymerase chain reaction; post-mortem genetic testing; sequencing

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