BCR-ABL融合基因定性检测试剂盒的性能评价  

Performance evaluation of BCR-ABL fusion gene qualitative testing kit

作　　者：李丽莉[1] 黄杰[1] 张文新 孙楠[1] 黄传峰[1] 曲守方[1] LI Lili;HUANG Jie;ZHANG Wenxin;SUN Nan;HUANG Chuanfeng;QU Shoufang(National Institutes for Food and Drug Control,Beijing,China,100050)

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《分子诊断与治疗杂志》2023年第8期1288-1291,共4页Journal of Molecular Diagnostics and Therapy

基　　金：国家重点研发计划诊疗装备与生物医用材料专项(2022YFC2409904)。

摘　　要：目的使用BCR-ABL参考品,评价BCR-ABL融合基因定性检测试剂盒的性能。方法提取参考品的RNA,测定其浓度和纯度。使用白血病融合基因检测试剂盒(荧光PCR法)进行PCR扩增。使用不同平台的荧光定量PCR仪进行检测,并使用仪器软件进行分析,获得参考品的BCR-ABL融合基因结果。结果阳性参考品在BCR-ABL反应通道扩增曲线有明显对数增长期且Ct值<试剂盒的阳性界值,为BCR-ABL融合突变阳性,BCR-ABL融合基因突变阴性和其他白血病融合基因突变的阴性参考品在BCR-ABL反应通道没有扩增曲线,为BCR-ABL融合突变阴性,不高于100拷贝/反应的检测限参考品在BCR-ABL反应通道扩增曲线有明显对数增长期且Ct值<阳性界值,为BCR-ABL融合突变阳性,重复性参考品的BCR-ABL反应通道Ct值的变异系数(CV,%)为0.4%~1.8%。结论BCR-ABL融合基因定性检测试剂盒能够准确检出参考品的BCR-ABL融合基因突变,符合《断裂点簇集区-艾贝尔逊白血病病毒(BCR-ABL)融合基因检测试剂盒》标准中的阳性参考品符合率、阴性参考品符合率、检出限和重复性项目的要求。Objective To evaluate the performance of BCR-ABL fusion gene qualitative testing kit with BCR-ABL reference.Methods The RNA of the BCR-ABL reference was extracted and its concentration and purity was measured.PCR was performed using the leukemia fusion gene testing kits(fluorescence PCR method).Fluorescence quantitative PCR instruments of different platforms were used for detection,and instrument software was used for analysis to obtain the BCR-ABL fusion gene result of the reference product.Results The positive reference product has a significant logarithmic growth phase in the amplification curve of the BCR-ABL reaction channel and the Ct value is<the positive cut-off value of the kit,which is positive for BCR-ABL fusion mutation.BCR-ABL fusion gene mutation negative and other leukemia fusion gene mutation negative reference products have no amplification curve in the BCR-ABL reaction channel,which is BCR-ABL fusion mutation negative.The detection limit reference product not higher than 100 copies/reaction has a significant logarithmic growth phase in the amplification curve of the BCR-ABL reaction channel and the Ct value is<the positive cut-off value,which is positive for the BCR-ABL fusion mutation.The coefficient of variation(CV,%)of the BCR-ABL reaction channel Ct value of the repeatable reference product was 0.4%~1.8%.Conclusion BCR-ABL fusion gene qualitative testing kit can accurately detect the BCR-ABL fusion gene mutation of the reference.Positive reference rate,negative reference rate,detection limit and reproducibility of the BCR-ABL fusion gene detection kit are satisfied with the requirements for industry standards of BCR-ABL fusion gene testing kit.

关 键 词：微小残留病 断裂点簇集区-艾贝尔逊白血病病毒融合基因 费城染色体 酪氨酸激酶抑制剂 

分 类 号：R733.7[医药卫生—肿瘤]