Abstract:

Objective To investigate the feasibility of detecting diffuse large B cell lymphoma（DLBCL） with the application of BIOMED-2 standardized immunoglobulin（IG） gene rearrangement detection in paraffin-embedded tissues. Methods DNA was extracted from 45 paraffin-embedded tissues. Multiple polymerase chain reaction（PCR） amplification was performed,and clonality analysis for IG gene rearrangement was performed by BIOMED-2 system primer. Results When DNA concentration was diluted to 50-100 ng/μL from 100-500 ng/μL,the proportion of the longest amplification fragment（300-400 bp） of DNA was improved from 10.0% to 90.0%. In 45 cases of DLBCL,the positive rate of immunoglobulin heavy chain（IGH）+ immunoglobulin kappa chain（IGK） was 84.4%. For IG/T-cell receptor（TCR） clonality analysis,no clonal rearrangement was found in 15 cases of reactive lymphoid tissue hyperplasia. Conclusions Dilution of DNA is a method to improve not only the proportion of the longest amplification fragment but also the detection rate of clonality. BIOMED-2 standardized immunoglobulin gene rearrangement detection is an efficient and reliable method for the diagnosis of DLBCL,with important clinical significance.

Key words: Gene rearrangement, Paraffin-embedded tissues, Diffuse large B cell lymphoma