TY - JOUR
T1 - DNA amplification from formalin-fixed decalcified paraffin-embedded bone marrow trephine specimens
T2 - Does the duration of storage matter?
AU - Talaulikar, Dipti
AU - Shadbolt, Bruce
AU - McNiven, Michelle
AU - Dahlstrom, Jane E.
PY - 2008/12
Y1 - 2008/12
N2 - Introduction: 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.
AB - Introduction: 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.
KW - Bone marrow trephines
KW - DNA
KW - Formalin-fixed paraffin-embedded
KW - Gene amplification
KW - Polymerase chain reaction (PCR)
UR - http://www.scopus.com/inward/record.url?scp=55549087905&partnerID=8YFLogxK
U2 - 10.1080/00313020802436410
DO - 10.1080/00313020802436410
M3 - Article
SN - 0031-3025
VL - 40
SP - 702
EP - 706
JO - Pathology
JF - Pathology
IS - 7
ER -