荧光定量PCR法检测弥漫大B细胞淋巴瘤IgH基因重排的意义

 目的 探讨荧光染料标记的实时定量PCR方法检测弥漫大B细胞淋巴瘤（DLBCL）IgH基因重排的可行性和临床意义。方法 44例DLBCL患者的57份骨髓标本用于检测IgH基因重排。Namalwa细胞系作阳性对照，U-937细胞系作阴性对照。SYBR Green荧光染料标记的实时荧光定量PCR方法检测IgH基因重排CDRⅢ。β-actin 作内参照，对IgH基因重排相对定量。结果 分析融解曲线可以确定IgH基因重排产物的特异性。荧光定量PCR检测IgH基因重排的阳性率分别为63.2 ％。Ⅰ、Ⅱ期患者IgH基因重排表达量中位数为0，Ⅲ、Ⅳ期患者IgH基因重排表达量中位数为0.35，两组患者之间差异有统计学意义（P＝0.018）。LDH值高于正常组，IgH基因重排表达量为0.39，LDH值低于正常组，IgH基因重排表达量为0.01，两组之间差异有统计学意义（P＝0.046）。结论 荧光染料标记的定量PCR方法可用于DLBCL的骨髓微小残留病变的检测。检测骨髓IgH基因重排，可以协助分期。

The significance of detecting IgH rearrangement in patients with diffuse large B-cell lymphoma using real-time flourescence quantitative PCR

Objective To investigate the feasibility and clinical significance of detecting IgH rear－rangement in diffuse large B cell lymphoma by SYBR Green RT-FQ-PCR. Methods Fifty-seven bone mar－row from 44 patients diagnosed with DLBCL were detected IgH-R. Namalwa cell line and U-937 cell line used for positive and negative control respectively. IgH-R CDR Ⅲ was detected by SYBR Green RT-FQ-PCR. The β-actin gene was chosen as inter control to normalize BM samples for DNA quantity. Results Melting curve analysis can confirm the specificity of IgH-R. The positive rate detected by RT-FQ-PCR was 63.2 %. 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 pa－tients with elevated LDH (P = 0.046). Conclusion SYBR Green RT-FQ-PCR is a valuable, feasible and sensi－tive tool to detect IgH rearrangement in DLBCL. Detecting IgH-R using RT-FQ-PCR can help staging.