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

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...

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

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 sites where the interpretation results of experimental group and control group were inconsistent,the verificaiton results was same as the experimental group. Conclusion The combined use of Triton X-100/4% PFA could improve the success rate of interphase FISH detection and the detection rate of positive sites in bone marrow samples.