Postmortem genetic testing for conventional autopsy-negative sudden unexplained death

An evaluation of different DNA extraction protocols and the feasibility of mutational analysis from archival paraffin-embedded heart tissue

Elisa Carturan, David J. Tester, Brian C. Brost, Cristina Basso, Gaetano Thiene, Michael John Ackerman

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

One third of autopsy-negative sudden unexplained deaths (SUDs) can be attributed to a cardiac channelopathy. Typically, paraffin-embedded tissue (PET) is the only source of DNA available for genetic analyses. We examined different DNA extraction procedures, involving 2 deparaffinization methods, 2 digestion methods, 4 laboratory-based purification methods, and 5 commercial kits. Mutational analysis involving 25 RYR2 exons was performed on PET DNA from 35 SUD cases to evaluate the feasibility of using PET DNA for genetic testing. With the best PET-DNA extraction method, an average of only two thirds of the region of interest could be evaluated. Although we initially identified 5 missense mutations in 5 of 35 SUD cases, repeated analysis failed to confirm these mutations. DNA from PET should be considered error prone and unreliable in comprehensive surveillance of SUD-associated genes. Given these shortcomings, the standard autopsy for SUD should include archiving EDTA-preserved blood or frozen tissue to facilitate postmortem genetic testing.

Original languageEnglish (US)
Pages (from-to)391-397
Number of pages7
JournalAmerican Journal of Clinical Pathology
Volume129
Issue number3
DOIs
StatePublished - Mar 2008

Fingerprint

Genetic Testing
Sudden Death
Paraffin
Autopsy
DNA
Channelopathies
Missense Mutation
Edetic Acid
Digestion
Exons
Mutation
Genes

Keywords

  • Autopsy
  • Catecholaminergic polymorphic ventricular tachycardia
  • DNA extraction
  • Genetic testing
  • Paraffin-embedded tissue
  • RYR2
  • Sudden unexplained death

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

@article{d33884c7bd6b4511b856c67f7e4c7de2,
title = "Postmortem genetic testing for conventional autopsy-negative sudden unexplained death: An evaluation of different DNA extraction protocols and the feasibility of mutational analysis from archival paraffin-embedded heart tissue",
abstract = "One third of autopsy-negative sudden unexplained deaths (SUDs) can be attributed to a cardiac channelopathy. Typically, paraffin-embedded tissue (PET) is the only source of DNA available for genetic analyses. We examined different DNA extraction procedures, involving 2 deparaffinization methods, 2 digestion methods, 4 laboratory-based purification methods, and 5 commercial kits. Mutational analysis involving 25 RYR2 exons was performed on PET DNA from 35 SUD cases to evaluate the feasibility of using PET DNA for genetic testing. With the best PET-DNA extraction method, an average of only two thirds of the region of interest could be evaluated. Although we initially identified 5 missense mutations in 5 of 35 SUD cases, repeated analysis failed to confirm these mutations. DNA from PET should be considered error prone and unreliable in comprehensive surveillance of SUD-associated genes. Given these shortcomings, the standard autopsy for SUD should include archiving EDTA-preserved blood or frozen tissue to facilitate postmortem genetic testing.",
keywords = "Autopsy, Catecholaminergic polymorphic ventricular tachycardia, DNA extraction, Genetic testing, Paraffin-embedded tissue, RYR2, Sudden unexplained death",
author = "Elisa Carturan and Tester, {David J.} and Brost, {Brian C.} and Cristina Basso and Gaetano Thiene and Ackerman, {Michael John}",
year = "2008",
month = "3",
doi = "10.1309/VLA7TT9EQ05FFVN4",
language = "English (US)",
volume = "129",
pages = "391--397",
journal = "American Journal of Clinical Pathology",
issn = "0002-9173",
publisher = "American Society of Clinical Pathologists",
number = "3",

}

TY - JOUR

T1 - Postmortem genetic testing for conventional autopsy-negative sudden unexplained death

T2 - An evaluation of different DNA extraction protocols and the feasibility of mutational analysis from archival paraffin-embedded heart tissue

AU - Carturan, Elisa

AU - Tester, David J.

AU - Brost, Brian C.

AU - Basso, Cristina

AU - Thiene, Gaetano

AU - Ackerman, Michael John

PY - 2008/3

Y1 - 2008/3

N2 - One third of autopsy-negative sudden unexplained deaths (SUDs) can be attributed to a cardiac channelopathy. Typically, paraffin-embedded tissue (PET) is the only source of DNA available for genetic analyses. We examined different DNA extraction procedures, involving 2 deparaffinization methods, 2 digestion methods, 4 laboratory-based purification methods, and 5 commercial kits. Mutational analysis involving 25 RYR2 exons was performed on PET DNA from 35 SUD cases to evaluate the feasibility of using PET DNA for genetic testing. With the best PET-DNA extraction method, an average of only two thirds of the region of interest could be evaluated. Although we initially identified 5 missense mutations in 5 of 35 SUD cases, repeated analysis failed to confirm these mutations. DNA from PET should be considered error prone and unreliable in comprehensive surveillance of SUD-associated genes. Given these shortcomings, the standard autopsy for SUD should include archiving EDTA-preserved blood or frozen tissue to facilitate postmortem genetic testing.

AB - One third of autopsy-negative sudden unexplained deaths (SUDs) can be attributed to a cardiac channelopathy. Typically, paraffin-embedded tissue (PET) is the only source of DNA available for genetic analyses. We examined different DNA extraction procedures, involving 2 deparaffinization methods, 2 digestion methods, 4 laboratory-based purification methods, and 5 commercial kits. Mutational analysis involving 25 RYR2 exons was performed on PET DNA from 35 SUD cases to evaluate the feasibility of using PET DNA for genetic testing. With the best PET-DNA extraction method, an average of only two thirds of the region of interest could be evaluated. Although we initially identified 5 missense mutations in 5 of 35 SUD cases, repeated analysis failed to confirm these mutations. DNA from PET should be considered error prone and unreliable in comprehensive surveillance of SUD-associated genes. Given these shortcomings, the standard autopsy for SUD should include archiving EDTA-preserved blood or frozen tissue to facilitate postmortem genetic testing.

KW - Autopsy

KW - Catecholaminergic polymorphic ventricular tachycardia

KW - DNA extraction

KW - Genetic testing

KW - Paraffin-embedded tissue

KW - RYR2

KW - Sudden unexplained death

UR - http://www.scopus.com/inward/record.url?scp=41649090413&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=41649090413&partnerID=8YFLogxK

U2 - 10.1309/VLA7TT9EQ05FFVN4

DO - 10.1309/VLA7TT9EQ05FFVN4

M3 - Article

VL - 129

SP - 391

EP - 397

JO - American Journal of Clinical Pathology

JF - American Journal of Clinical Pathology

SN - 0002-9173

IS - 3

ER -