CAJ | 학술논문

目的探讨联合免疫荧光和荧光原位杂交(FISH)的FICTION技术在多发性骨髓瘤(MM)遗传学异常检测中的应用价值.方法采集18例MM患者和2例浆细胞白血病患者的骨髓标本,分离单个核细胞制作滴片.从细菌人工染色体文库中选取覆盖IgH、MMSET待测基因位点的质粒,用缺口平移法制备带有半抗原检测标签的核酸探针.在经CD138标记和酪胺信号放大的细胞滴片标本上,使用上述自制探针[t(4;14)、t(11;14)和t(14;16)]和商品化直标缺失探针(13q和p53)进行FISH检测.结果 20例患者标本均使用上述5种探针进行检测,其中检出t(4;14)4例,t(11;14)6例,t(14;16)1例,p53缺失3例,13q缺失8例;另有4例未检测出此5种异常.结论应用FICTION技术原位分析骨髓中特定瘤细胞亚群的特征性遗传学异常,能够提高FISH检测的效率和敏感性,并可作为对MM患者遗传学分层诊断的初筛实验,指导治疗并判断预后.
목적 탐토연합면역형광화형광원위잡교(FISH)적FICTION기술재다발성골수류(MM)유전학이상검측중적응용개치.방법 채집18례MM환자화2례장세포백혈병환자적골수표본,분리단개핵세포제작적편.종세균인공염색체문고중선취복개IgH、MMSET대측기인위점적질립,용결구평이법제비대유반항원검측표첨적핵산탐침.재경CD138표기화락알신호방대적세포적편표본상,사용상술자제탐침[t(4;14)、t(11;14)화t(14;16)]화상품화직표결실탐침(13q화p53)진행FISH검측.결과 20례환자표본균사용상술5충탐침진행검측,기중검출t(4;14)4례,t(11;14)6례,t(14;16)1례,p53결실3례,13q결실8례;령유4례미검측출차5충이상.결론 응용FICTION기술원위분석골수중특정류세포아군적특정성유전학이상,능구제고FISH검측적효솔화민감성,병가작위대MM환자유전학분층진단적초사실험,지도치료병판단예후.
Objective To investigate the diagnostic value of FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms) technique, combining immunofluorescence and fluorescence in situ hybridization (FISH), to detect genetic aberrations in multiple myeloma (MM). Methods Bone marrow samples were collected from 18 MM and 2 plasma cell leukemia (PCL)patients. Probes targeting IgH and MMSET were prepared using a Nick Translation Kit from Bacterial artificial chromosome (BAC) clones. The immunophenotyping was achieved via the CD138 tyramide signal amplification (TSA)-mediated immunofluorescence, followed by FISH with the prepared probes [t (4;14), t (11;14), t(14;16)] and the commercial deletion probes (13q and p53) to detect common genetic aberrations in MM. Results All the 20 samples were assayed with the probes mentioned above, and revealed 4 cases with t(4;14) ,6 with t(11 ;14), 1 with t(14;16), 3 with p53 deletion; and 8 with 13q deletion. The remaining 4 cases had none of the 5 aberrations. Conclusion FICTION technique facilitates the detection of genetic abnormalities of MM in situ; enhances both efficiency and sensitivity of positive det~tion, thus, could be used as the screening test of molecular diagnosis of MM to guide coming-up risk-adapted therapy and evaluate prognosis.