Objective: To explore the application of BIOMED- 2 standardized immunoglobulin (Ig) gene rearrangement system in the diagnosis of multiple myeloma (MM), and the significance of clonality analysis by multiplex-PCR amplifications.
Methods: A total of 167 cases of MM bone marrow samples from 2009 to 2013, and 20 cases of reactive plasmacytosis used as the controls were included in this study. Multiplex-PCR amplifications were performed and the Ig gene rearrangements were analyzed using BIOMED-2 standardized clonality analysis system.
Results: ① Of 167 MM cases, 107 showed IgH VH-JH rearrangement, 33 showed IgH DH-JH rearrangement, and 30% showed IgH DH-JH rearrangement in 60 IgH VH-JH rearrangement negative MM cases. The difference was statistically significant between IgH VH-JH rearrangement positive and negative cases (14.0% vs 30.0%, P=0.032). The total positive rate of IgH VH-JH, IgH DH-JH and IgK was 94.6%. The 20 reactive plasmacytosis (RP) cases showed negative Ig gene rearrangement. 2 of 167 MM cases, 9 (5.4%) showed clonal IgH rearrangement by agarose electrophoresis were confirmed as polyclonality by capillary electrophoresis. ③ Of 53 MM cases who have been detected by Ig gene rearrangement system and fluorescence in situ hybridization (FISH) for IgH simultaneously, 36 showed IgH rearrangement, 26 showed FISH IgH positive, and the difference was statistically significant (67.9% vs 49.1%, P=0.049).
Conclusion: Combined detection of IgH VH- JH, IgH DH- JH and IgK could improve the positive rate of MM clonality dramatically, and measurement of IgH DH-JH rearrangement was more important in the IgH VH- JH negative cases. Ig gene rearrangement system was a faster and more sensitive method than FISH IgH. Application of BIOMED- 2 standardized immunoglobulin (Ig) gene rearrangement system is of significance for MM diagnosis.
目的: 探讨BIOMED-2标准化Ig基因重排技术在多发性骨髓瘤（MM）诊治中的应用以及采用PCR片段分析毛细管电泳法（简称毛细管电泳法）进行克隆性检测的意义。
方法: 选取2009年至2013年167例MM患者骨髓标本，以20例反应性浆细胞增多症患者骨髓标本作为对照，采用BIOMED-2系统引物，进行多重PCR扩增并采用毛细管电泳法进行Ig基因重排的克隆性分析。
结果: ①167例MM患者中IgH VH-JH重排阳性者107例，阴性者60例，两组患者中IgH DH-JH重排发生率差异有统计学意义（14.0％对30.0%，P=0.013）；121例（72.4%）患者存在IgK重排。IgH VH-JH、IgH DH-JH、IgK联合检查，阳性检出率为94.6%。对照组Ig基因重排检测结果均为阴性。②琼脂糖电泳法结果示167例MM患者中9例（5.4%）患者IgH重排基因克隆性阳性，而毛细管电泳法结果则示为多克隆重排。③53例MM患者同时进行Ig基因重排检测和染色体荧光原位杂交IgH检测，两种方法的IgH阳性检出率差异有统计学意义（67.9％对49.1%，P=0.049）。
结论: IgH VH-JH、IgH DH-JH、IgK联合检测可大大提高MM患者Ig基因重排克隆性的检出率，尤其对于IgH VH-JH重排阴性者IgH DH-JH重排的补充检测尤为重要。基因重排检测采用毛细管电泳法可有效地区分单克隆、多克隆和寡克隆。Ig基因重排检测较染色体荧光原位杂交技术的IgH阳性检出率高。应用BIOMED-2标准化Ig基因重排技术检测对于MM的诊断有指导意义。