BIOMED-2 standardized immunoglobulin gene rearrangement detection in paraffin-embedded tissues for diffuse large B cell lymphoma

doi:10.3969/j.issn.1673-8640.2016.09.017

Abstract

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

CLC Number:

R446.1

SUN Juan, AI Xiaofei, LI Qinghua, CAO Zeng. BIOMED-2 standardized immunoglobulin gene rearrangement detection in paraffin-embedded tissues for diffuse large B cell lymphoma[J]. Laboratory Medicine, 2016, 31(9): 810-813.

Figures/Tables 2

References 8

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PCR产物最大片段（bp）	DNA标本原浓度（30例）		原浓度DNA标本稀释到50~100 ng/μL（30例）
PCR产物最大片段（bp）	100~300 ng/μL	300~500 ng/μL	原浓度DNA标本稀释到50~100 ng/μL（30例）
100	6（20.0）	10（33.3）	0（0.0）
200	7（23.3）	4（13.3）	3（10.0）
300	2（6.7）^*	1（3.3）^*	26（86.7）^*
400	0（0.0）	0（0.0）	1（3.3）

PCR产物最大片段（bp）	DNA标本原浓度（30例）		原浓度DNA标本稀释到50~100 ng/μL（30例）
PCR产物最大片段（bp）	100~300 ng/μL	300~500 ng/μL	原浓度DNA标本稀释到50~100 ng/μL（30例）
100	6（20.0）	10（33.3）	0（0.0）
200	7（23.3）	4（13.3）	3（10.0）
300	2（6.7）^*	1（3.3）^*	26（86.7）^*
400	0（0.0）	0（0.0）	1（3.3）

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