摘要
本研究旨在明确免疫磁珠浆细胞分选对使用荧光原位杂交(FISH)技术检测多发性骨髓瘤(MM)细胞分子遗传学异常的影响,探索适合我国国情的检测方法。对同一MM患者的标本进行2种方法检测:1种为不经分选,直接进行FISH;另1种为经过CD138免疫磁珠分选(MACS)富集浆细胞后进行FISH检测;比较2种FISH技术对MM遗传学异常检出率并分析其影响因素。探针类型为13q14(RB-1)和14q32(IGH)。结果表明:①利用13q14(RB-1)探针对42例患者同时进行了分选和不分选的的FISH检测显示,经过CD138磁珠分选后25例(56.8%)检测到13q14缺失;不经分选直接进行FISH检测仅9例(21.4%)患者检出13q14缺失,2种标本处理方法的检出率具有统计学差异(p=0.01);②利用14q32(IGH)探针对22例患者同时进行了分选和不分选的FISH检测显示,经过CD138磁珠分选后14例(63.6%)可以检测到14q32重排,不经分选直接进行FISH检测仅7例(31.8%)患者检出14q32重排,2种标本处理方法的检出率具有统计学差异(p=0.035);③不分选FISH的阳性细胞检出率与骨髓涂片和流式细胞仪所检测的浆细胞比例呈明显的正相关。当骨髓涂片细胞中浆细胞比例超过50%,或者流式细胞仪检测浆细胞比例超过10%时,不分选FISH与磁珠分选FISH结果之间没有统计学差异。结论:MACS-FISH显著提高了MM遗传学异常的检出率,避免了假阴性的发生;直接进行FISH检测的检出率低的主要原因是检测标本中浆细胞比例较低;未经分选直接进行13q14(RB-1)和14q32(IGH)探针FISH检测,其阳性细胞率与骨髓涂片、流式细胞仪所检测出的浆细胞比例呈显著正相关,说明随着所检验标本中浆细胞比例的升高,不进行分选直接进行FISH检测的缺点得到一定弥补。当骨髓涂片细胞中浆细胞比例超过50%,或者流式细胞仪检测浆细胞比例超过10%时,2种检测方法之间没有差异。
This study was to aimed investigate the influence of immunomagnetic sorting on detecting the genetic aberrations of multiple myeloma（MM） by interphase fluorescence in situ hybridization（FISH） and to explore the detection method suitable to use in our country.The genetic aberrations of immunomagnetically sorted and unsorted bone marrow cells from the same MM patients were detected by interphase FISH and the detectable rate of genetic aberration was compared.The types of probes included 13 q14（RB-1） and 14q32（IGH）.The 42 and 22 sorted and unsorted marrow samples from MM patients were detected by using 13q14 probe and 14q32 probes respectively,the results indicated that the 13q14 deletion was found in 9 of 42（21.4%） unsorted marrow samples and in 25 of 42（56.8%） CD138＋-sorted marrow samples.The 13q32 rearrangement was found in 7 of 22（31.8%） unsorted marrow samples and in 14 of 22（63.6%） CD138＋-sorted marrow samples.Both of the difference was statistically significant（p=0.001 and p=0.035 respectively）.Percentages of cytogenetic alterations detected in unsorted bone marrow cells correlated positively with percentage of plasma cells tested by bone marrow smears or flow cytometry.When percentage of plasma cells tested by bone marrow smears exceed 50%,or by flow cytometry exceed 10%,there was no difference between 2 methods.It is concluded that immunomagnetic sorting of CD138＋ cells increases the probability of detection of the 13q14 deletion and 14q32 rearrangement in bone marrow samples.The low detectable rate of genetic aberration in unsor-ted bone marrow cells is associated to the low percentage of plasma cells in bone marrow samples,higher percentage of plasma cells can partly overcome the shortage of unsorted detection method.When percentage of plasma cells tested by bone marrow smears exceed 50%,or by flow cytometry exceed 10%,there was no difference between 2 methods.
出处
《中国实验血液学杂志》
CAS
CSCD
2011年第1期54-58,共5页
Journal of Experimental Hematology
基金
天津市科技支撑计划项目(编号09ZCGYSF1000)
天津市科技计划项目(编号09ZCZDSF3800)
卫生部临床科学重点项目基金
关键词
多发性骨髓瘤
荧光原位杂交
免疫磁珠分选
multiple myeloma
fluorescence in situ hybridization
immunomagnetic sorting
