The study of immunoglobulin heavy-chain gene rearrangement in diffuse large B-cell lymphoma using fluorescence quantitative PCR
- VernacularTitle:荧光定量PCR分析弥漫大B细胞淋巴瘤免疫球蛋白重链基因重排表达
- Author:
Jiqun YI
;
Tongyu LIN
;
Youjian HE
- Publication Type:Journal Article
- Keywords:
diffuse large B cell lymphoma;
immunoglubulin heavy chain gene;
real-time fluorescence quantitative PCR;
nested PCR
- From:
China Oncology
2006;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Background and purpose:Majority of patients with B cell lymphoma often achieve complete clinical remission after systemic treatment, but half of the patients ultimately relapse. The residual neoplastic cells, commonly called ‘minimal residual disease’ (MRD), are thought to be the source of relapse. But not all of the patients whose results of IgH rearrangement were positive had relapse or distant involvement. It was thought that the patients whose IgH rearrangement was positive relapsed or not may be associated with the quantity of IgH rearrangement. The study tried to investigate the feasibility and clinical significance of detecting immunoglobulin heavy chain gene in DLBCL by SYBR Green RT-FQ-PCR. Methods:Fifty-seven bone marrow specimens from 44 patients diagnosed with DLBCL were used to detect IgH-R. Namalwa cell line and U-937 cell line were used for positive and negative control respectively. The ?-actin gene was chosen as inter control. DNA was isolated by phenol-chloroform-isoamyl alcohol and then was amplified by SYBR Green RT-FQ-PCR targeting the IgH-R CDR Ⅲ.Results:Melting curve analysis could confirm the specificity of IgH-R. The positive rate detected by RT-FQ-PCR was 63.2%. The positive results of IgH/?-actin were between 0.01 and 4131.69, and the median was 0.42. There was a significant difference between stage Ⅰ/Ⅱ and stage Ⅲ/Ⅳ in IgH-R quantity (P=0.018). Nonparametric test showed that there was a significant difference between patients with normal LDH and patients with elevated LDH (P=0.046).Conclusions:SYBR Green RT-FQ-PCR is a valuable, feasible and sensitive tool to detect IgH rearrangement in DLBCL. Detecting IgH-R using RT-FQ-PCR can help staging more accurately.
