应用多重PCR技术检测慢性淋巴细胞白血病IgVH基因突变  

Detection of IgVH Mutation Status in Patients with Chronic Lymphocytic Leukemia by Multiplex PCR

作　　者：郑文娟[1] 陈丽娟[1] 吴雨洁[1] 李丽[1] 徐卫[1] 李建勇[1] 

机构地区：[1]南京医科大学第一附属医院,江苏省人民医院血液科,南京210029

出　　处：《中国实验血液学杂志》2006年第6期1101-1104,共4页Journal of Experimental Hematology

基　　金：江苏省"135工程"医学重点人才基金资助;编号RC2002044

摘　　要：免疫球蛋白重链可变区(IgVH)基因突变是慢性淋巴细胞白血病(CLL)最重要的独立预后因素之一,为探讨CLL患者IgVH基因突变状态,应用多重PCR技术检测9例CLL患者的IgVH基因,纯化PCR扩增产物后直接测序,应用IMGT/V-QUEST工具分析,明确有无IgVH突变及突变位置。结果表明9例CLL患者皆扩增出395-465bp区域(MIXI)或290-360bp区域(MIXII)单克隆条带,测序结果显示5例患者有突变,IgVH基因分别为IGHV3-1103、IGHV3-901、IGHV3-2301、IGHV4-5901和IGHV4-3402;4例无突变,IgVH基因为IGHV3-5301、IGHV3-2303、IGHV3-3305和IGHV3-701。结论PCR检测IgVH基因突变,简化了繁琐的实验过程,缩短了实验时间,解决传统PCR对IgVH基因突变检测的桎梏,值得在临床和科研中推广使用。IgVH mutation status is one of the most important independent prognostic factor of chronic lymphocytic leukemia （CLL）. In order to evaluate IgVH mutation status in patients with CLL, IgVH mutation was detected by multiplex PCR in 9 CLL patients and purified PCR amplification products were directly sequenced, IgH somatic hypermutation and mutation site were analysed by IMGT/V-QUEST. The results showed that 5 patients had mutated IgVH, and IgVHs were IGHV3-11 ＊ 03, IGHV3-9 ＊ 01, IGHV3-23 ＊ 01, IGHV4-59 ＊ 01, IGHV4-34 ＊ 02, respectively ; whereas 4 others had unmutated IgVH, these IgVHs were IGHV3-53 ＊ 01, IGHV3-23 ＊ 03, IGHV3-33 ＊ 05 and IGHV3-7 ＊ 01. It is concluded that multiplex PCR is a rapid and easy method to detect IgVH mutation status, and it solves the limitations and pitfalls of routine PCR, and it is worth being extensively used both in clinic and scientific researches.

关 键 词：多重PCR IGVH 基因突变 

分 类 号：R733.72[医药卫生—肿瘤]