联合运用Triton X-100和多聚甲醛改善血液肿瘤骨髓标本荧光原位杂交检测质量  

Combining Triton X-100 and paraformaldehyde to optimize quality of bone marrow FISH detection of hematological tumors

作　　者：傅蔷 谢珊珍 叶常姜 黄慧芳[2] FU Qiang;XIE Shanzhen;YE Changjiang;HUANG Huifang(Fujian Medical University Union Hospital,ujian Provincial Key Laboratory on Hematology,Central Laboratory,Fuzhou 350001,Fujian;Fujian Institute of Hematology,Fujian Provincial Key Laboratory on Hematology,b.Central Laboratory,Fuzhou 350001,Fujian;Beijing GP Medical Technologies,Ltd.Beijing 100176,China)

机构地区：[1]福建医科大学附属协和医院福建省血液病研究所,福建省血液病学重点实验室,福州350001 [2]福建医科大学附属协和医院中心实验室,福州350001 [3]北京金菩嘉医疗科技有限公司,北京100176

出　　处：《临床检验杂志》2021年第1期26-29,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家临床重点专科建设项目;福建省临床重点专科建设项目;福建省血液医学中心建设项目[闽政办(2017)4号];福建省高水平实验研究平台建设项目(闽201704);福建医科大学启航基金(2016QH031)。

摘　　要：目的通过联合运用Triton X-100和多聚甲醛(PFA)处理玻片,改善血液肿瘤骨髓标本荧光原位杂交(FISH)检测质量。方法42例血液肿瘤患者骨髓标本玻片,在常规FISH流程中,实验组增加Triton X-100/4%PFA预处理步骤,对照组按照常规步骤进行。配对比较实验组与对照组中未见杂交信号细胞数和阳性细胞比例。当实验组与对照组结果不一致时,骨髓增生异常综合征(MDS)标本通过扩大计数范围验证准确性,多发性骨髓瘤(MM)标本采用荧光免疫表型结合间期原位杂交(FICTION)检测验证准确性。结果与对照组相比,实验组杂交区域非特异性荧光信号减少,背景干净,细胞核内探针信号更清晰,FISH检测成功率达100%。实验组中未见杂交信号的细胞比例为(4.30±3.30)%,对照组为(8.04±5.45)%,二者配对差值(实验组-对照组)为(-3.74±4.27)%,实验组未见杂交信号的细胞比例低于对照组(P<0.001)。实验组与对照组判读结果一致的132个信号位点中,实验组的阳性细胞比例为(82.16±23.89)%,对照组为(77.10±23.56)%,二者配对差值(实验组-对照组)为(5.06±10.08)%,实验组阳性细胞比例高于对照组(P<0.001)。此外,实验组与对照组的判读结果不一致的5个检测位点,验证结果与实验组结果一致。结论联合运用Triton X-100/4%PFA能够提高骨髓标本间期FISH检测成功率和阳性位点检出率。Objective To improve the quality of fluorescence in situ hybridization( FISH) detection of hematological tumor bone marrow samples bycombination of Triton X-100 and paraformaldehyde( PFA) in pretreatment of sample slides. Methods In routine FISH procedure for the bone marrow specimen slides from 42 patients with hematological tumors,the experimental group added Triton X-100/4% PFA in pretreatment step,and the control group followed the routine steps. The number of cells without hybridization signal and the proportion of positive cells in both experimental group and control group were paired and compared. When the results of experimental group and control group were inconsistent,the accuracy of the myelodysplastic syndrome( MDS) samples was verified by expanding the count range and the accuracy of multiple myeloma( MM) samples was detected and verified by fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasms( FICTION). Results Compared with the control group,the experimental group showed fewer non-specific fluorescence signals in the hybridization area,clean background and clearer probe signals in the nucleus with success rate of 100% in FISH detection. In experimental group,the average of the proportion of the cells without hybridization signal was( 4.30±3.30) %,while it was( 8.04±5.45) % in control group,and the paired difference between the experimental and control group was(-3.74±4.27) %. In the experimental group the proportion of cells without hybridization signal was signifiantly lower than that in control group( P<0.001). Among the 132 signal sites where the interpretation results of experimental group and control group were consistent,the average ratio of positive cells in experimental group was( 82.16±23.89) %,and( 77.10± 23.56) % in control group. The paired difference between the two groups was( 5.06±10.08) %,and the proportion of positive cells in experimental group was significantly higher than that in control group( P<0.001). In the 5 detection sit

关 键 词：荧光原位杂交 Triton X-100 多聚甲醛 骨髓 

分 类 号：R446[医药卫生—诊断学]