[Rapid DNA extraction from archival paraffin-embedded tissue: a modified method available to PCR analysis for diagnostic pathology].
OBJECTIVE
To modify a method for rapid DNA extraction from paraffin-embedded tissue and to evaluate its efficiency and feasibility available to the PCR analysis for the diagnostic pathology.


METHODS
Rapid DNA extraction by simply boiling was performed on 52 archival paraffin-embedded tissues including lymphoma and reactive hyperplasia. The traditional phenol-chloroform and DNeasy mini kit methods were also used as controls. The DNA quality was confirmed by PCR amplification of housekeeping gene beta-Globin. The accuracy of target fragment was reconfirmed by sequencing analysis. Based on the clinical pathologic diagnosis to diseases, the IgH and TCRY gene were selected for their gene rearrangements detected.


RESULTS
The amplification efficiency of beta-Globin and the detective rate of IgH and TCRY gene rearrangements showed no statistic significance between rapid method and the controls, although an obvious lower DNA concentration extracted by rapid method was showed by spectrophotometer. The sequencing result of PCR product was thoroughly consistent with the DNA sequence of corresponding target gene. The DNA in extract could be stored at -20 degrees C for at least 4 weeks.


CONCLUSIONS
The rapid method of DNA extracted from archival paraffin-embedded tissue can provide enough DNA served as PCR template, and be regarded as a reliable, stable and economical method with prospect in being used to PCR analysis for the diagnostic pathology.
Jiang
Wei
W
Department of Pathology, West China Hospital, Sichuan University, Chengdu 610041, China.
Liu
Wei-ping
WP
Li
Lei
L
Li
Gan-di
GD
D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D003933:Diagnosis; D004586:Electrophoresis; D005240:Feasibility Studies; D006801:Humans; D008223:Lymphoma / Q000175:diagnosis / Q000235:genetics / Q000473:pathology; D016612:Paraffin Embedding / Q000379:methods*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D013048:Specimen Handling; D013997:Time Factors
D004247:DNA