Specimen stability for DNA-based diagnostic testing.
The use of molecular diagnostic testing is increasing in the clinical setting; therefore, data regarding DNA stability in clinical specimens are essential for correct test performance and interpretation. This study was designed to determine DNA stability in peripheral blood and solid tissue under different storage conditions. DNA quality and yield were assayed by spectrophotometric absorbance, gel electrophoresis, and suitability for Southern hybridization and polymerase chain reaction (PCR), the most widely employed clinical DNA analyses. A second goal of the study was to evaluate DNA stability during storage at 4 degrees C for 1 month to 3 years. The data show that freezing or refrigeration of separated leukocytes is preferable for short- to intermediate-term storage and freezing is preferable for solid tissue. DNA degradation varying from slight to severe is seen inconsistently with such specimens, probably due to sampling of unevenly frozen-tissue areas. Depending on the degree of DNA degradation, analysis may still be possible by PCR and in some cases even by Southern hybridization. Once isolated, DNA was stable at 4 degrees C for at least 3 years. These results suggest a more flexible approach to specimen requirements for molecular pathology, as some samples that would routinely be rejected gave interpretable results.
Farkas
D H
DH
Department of Clinical Pathology, William Beaumont Hospital, Royal Oak, Michigan 48073-6769, USA.
Drevon
A M
AM
Kiechle
F L
FL
DiCarlo
R G
RG
Heath
E M
EM
Crisan
D
D
D001773:Blood Cells / Q000648:ultrastructure; D015139:Blotting, Southern; D003582:Cytogenetics; D004247:DNA / Q000032:analysis* / Q000302:isolation & purification; D004262:DNA Restriction Enzymes; D004586:Electrophoresis; D005821:Genetic Techniques; D006801:Humans; D010920:Placenta / Q000648:ultrastructure; D016133:Polymerase Chain Reaction; D013048:Specimen Handling; D013696:Temperature; D014021:Tissue Preservation
D004247:DNA; D004262:DNA Restriction Enzymes