DNA amplification from formalin-fixed decalcified paraffin-embedded bone marrow trephine specimens: does the duration of storage matter?
BACKGROUND
Examination of bone marrow (BM) trephines has been traditionally limited to histological analysis. With renewed interest in their use in molecular diagnostics, studies assessing the impact of duration of storage on DNA amplification from archived BM trephines are required.


METHODS
DNA extraction was performed on 169 formalin-fixed decalcified paraffin-embedded (FFDPE) trephine blocks with a mean duration of storage of 59.4 months (range 4-200 months). An amplification control master mix from the In Vivo Scribe Immunoglobulin Heavy Chain Clonality kit was used to determine DNA amplification at different amplicon sizes [96-600 base pairs (bp)].


RESULTS
Amplification at 96 bp was noted in 145 of 169 specimens (85.8%). Progressive reduction in amplification was noted at higher base pairs with 99 of 169 (58.6%) amplifying at 200 bp, 56 cases (33.1%) at 300 bp, 35 (20.7%) at 400 bp and 19 (11.2%) at 600 bp. The impact of duration of storage on DNA amplification was not statistically significant.


CONCLUSIONS
DNA was obtained from archival BM trephine specimens in approximately 86% of cases, indicating good potential for utilisation in polymerase chain reaction (PCR) techniques. The duration of storage of FFDPE tissue had no impact on the ability to amplify specimens.
Talaulikar
Dipti
D
Department of Haematology, The Canberra Hospital, Yamba Drive, Garran, Canberra, ACT 2606, Australia. dipti.talaulikar@act.gov.au
Shadbolt
Bruce
B
McNiven
Michelle
M
Dahlstrom
Jane E
JE
D001706:Biopsy; D001853:Bone Marrow / Q000473:pathology*; D004247:DNA / Q000032:analysis*; D003648:Decalcification Technique; D005557:Formaldehyde; D006801:Humans; D008232:Lymphoproliferative Disorders / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D016612:Paraffin Embedding; D013048:Specimen Handling / Q000379:methods*; D013995:Time; D016707:Tissue Fixation
D005557:Formaldehyde; D004247:DNA