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CD138免疫磁珠分选骨髓瘤细胞联合荧光原位杂交技术在检测多发性骨髓瘤细胞遗传学异常中的应用

[Application of CD138 Immunomagnetic Sorting Myeloma Cells Combined with Fluorescence in Situ Hybridization for Detecting Cytogenetic Abnormalities of Multiple Myeloma].

作者信息

Gao Lu, Liu Qing, Shi Yan, Dang Hui, He Qi, Wang Zheng, Feng Lin, Li Ye, Wang Xiao-Yan, Li Na, Song Wen-Jie, Wang Yan-Lin, Kong Shu, Lu Jin, Huang Xiao-Jun, Lai Yue-Yun

机构信息

Beijing Key Laboratory of Hematology Stem Cell Transplantation, Peking University People's Hospital, Peking University Institute of Hematology, Beijing 100044,China.

Beijing Key Laboratory of Hematology Stem Cell Transplantation, Peking University People's Hospital, Peking University Institute of Hematology, Beijing 100044,China. E-mail:

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Jun;25(3):807-812. doi: 10.7534/j.issn.1009-2137.2017.03.031.

Abstract

OBJECTIVE

To investigate the efficiency of direct fluorescence in situ hybridization (D-FISH) versus FISH on CD138 immunomagnetic sorting myeloma cells (MACS-FISH) to detect the cytogenetic abnormalities of multiple myeloma.

METHODS

Thirty-one patients with multiple myeloma (MM) were detected by D-FISH and MACS-FISH, using 5 probes, including 1q21, D13S319, RB1, IgH, P53. The IgH rearrangement positive patients were further examined by 3 IgH rearrangement subtype FISH probes including IgH/FGFR3, IgH/MAF and IgH/CCND1.

RESULTS

Metaphase karyotyping revealed cytogenetic abnormalities in 5 cases (16.1%), clonal aberrations were detected in 13 cases(41.9%) by D-FISH, while 25 case(80.6%) with clonal aberrations by MACS-FISH. The results between these 2 FISH methods were significantly different (P=0.042). The detection frequency of clonal aberration by each probes of D-FISH was 22.6%,25.8%,29%,38.7% and 9.7% respectively for 1q21 amplification, D13S319 deletion,RB1 deletion, IgH rearrangement and P53 deletion, compared with 48.4%,45.2%,48.4%,67.7% and 16.1% respectively by MACS-FISH. The 2 FISH methods were well consistent when the percentage of plasma cells was ≥20% in bone marrow smears. When the percentage of plasma cells was<20% in bone marrow smears, the difference between these 2 methods was very statistically significant (P=0.00).

CONCLUSION

MACS-FISH can obviously improve the detection efficiency of cytogenetic abnormalities in patients with MM. Conventional cytogenetics combined with MACS-FISH is an ideal efficient method to detect the cytogenetic abnormalities in MM patients, and should be applied widely, especially for those patients with the plasma cells <20% in bone marrow smears.

摘要

目的

探讨直接荧光原位杂交(D-FISH)与CD138免疫磁珠分选骨髓瘤细胞荧光原位杂交(MACS-FISH)检测多发性骨髓瘤细胞遗传学异常的效率。

方法

采用包括1q21、D13S319、RB1、IgH、P53在内的5种探针,对31例多发性骨髓瘤(MM)患者进行D-FISH和MACS-FISH检测。对IgH重排阳性患者,进一步采用包括IgH/FGFR3、IgH/MAF和IgH/CCND1在内的3种IgH重排亚型FISH探针进行检测。

结果

中期核型分析显示5例(16.1%)存在细胞遗传学异常,D-FISH检测到13例(41.9%)存在克隆性畸变,而MACS-FISH检测到25例(8�.6%)存在克隆性畸变。这两种FISH方法的结果差异有统计学意义(P=0.042)。D-FISH各探针检测克隆性畸变的频率分别为1q21扩增22.6%、D13S319缺失25.8%、RB1缺失29%、IgH重排38.7%、P53缺失9.7%,而MACS-FISH分别为48.4%、45.2%、48.4%、67.7%和16.1%。当骨髓涂片浆细胞比例≥20%时,两种FISH方法结果一致性良好。当骨髓涂片浆细胞比例<20%时,两种方法差异有极显著统计学意义(P=0.00)。

结论

MACS-FISH可明显提高MM患者细胞遗传学异常的检测效率。传统细胞遗传学联合MACS-FISH是检测MM患者细胞遗传学异常的理想高效方法,应广泛应用,尤其适用于骨髓涂片浆细胞比例<20%的患者。

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