BCR::ABL1 p210转录水平定量检测多中心比对

Comparison of quantitative detection of BCR::ABL1 p210 transcript levels:a multicenter study

目的评估7家独立医学实验室BCR::ABL1 p210转录水平定量检测能力及实验室之间结果的可比性。方法比对分两个阶段完成,由参考实验室制备比对样本,每个阶段比对样本的BCR::ABL1 p210定量值均覆盖0.001%~0.01%、0.01%~0.1%、0.1%~1%、1%~10%及>10%。各比对实验室分别采用即时荧光定量PCR法(real-time quantitative PCR,RT-PCR)和数字PCR法(digital PCR,dPCR)进行比对样本的检测,计算并确认各比对实验室转换因子(conversion factor,CF)。结果(1)RT-PCR比对中,1家实验室第一阶段14例样本未检出,其余6家实验室比对结果合格,偏倚<±1.2倍(-0.133~0.338),95%一致性界限<±5倍(上限0.147~0.785,下限-0.770~-0.109),计算并确认CF值;(2)dPCR比对中,1家实验室第二阶段未回报结果,其余6家实验室比对结果合格,偏倚<±1.2倍(-0.026~0.267),95%一致性界限<±5倍(上限0.084~0.991,下限-0.669~-0.135),计算并确认CF值;(3)BCR::ABL1 p210定量值为0.01%~0.1%、0.1%~1%、1%~10%及>10%时,RT-PCR和dPCR均能全部检出;BCR::ABL1 p210定量值为0.001%~0.01%时,dPCR检出率高于RT-PCR法(85.56%比68.00%)。结论各比对实验室结果呈较好一致性,各实验室间BCR::ABL1 p210定量检测通过CF值换算具备可比性;BCR::ABL1 p210深度分子反应定量检测中,dPCR法阳性检出率更高,更有优势;为保证检测结果的准确性和稳定性,各实验室应加强日常质控工作。

Objective To assess the capability of seven reference medical laboratories to detect BCR::ABL1 p210 transcription levels and to compare the results among those laboratories.Methods The interlaboratory comparison was carried out in two stages.The samples were prepared by the reference laboratory.The quantitative values of BCR::ABL1 p210 of the comparison samples covered 0.001%-0.01%,0.01%-0.1%,0.1%-1%,1%-10%and>10%in each stage.Real-time quantitative PCR(RT-PCR)and dPCR(digital PCR)were used to examine the samples.The conversion factor(CF)was calculated and validated for each laboratory.Results In the RT-PCR comparison,one laboratory was failed to detect BCR::ABL1 p210 in fourteen samples at the first stage.The results of the other six laboratories were qualified with the bias<±1.2 folds(-0.133-0.338)and 95%limits of agreement within±5 folds(upper limit 0.147-0.785,lower limit-0.770--0.109),and the corresponding CF values were calculated and validated.In the dPCR comparison,one laboratory did not report results at the second stage.The results of the other six laboratories were qualified with the bias<±1.2 folds(-0.026-0.267)and 95%limits of agreement within±5 folds(upper limit 0.084-0.991,lower limit-0.669--0.135),and the corresponding CF values were calculated and validated.The samples with BCR::ABL1 p210 quantitative values of 0.01%-0.1%,0.1%-1%,1%-10%and>10%could be detected by both RT-PCR and qPCR.When the quantitative value of BCR::ABL1 p210 was 0.001%-0.01%,the detection rate of dPCR was higher than that of RT-PCR(85.56%vs.68.00%).Conclusions A good consistency is present among various laboratories.The quantitative value of BCR::ABL1 p210 is comparable among laboratories as shown by the CF value conversion.For quantitative detection of BCR::ABL1 p210 deep molecular reaction,dPCR has a higher positive detection rate and more advantages than RT-PCR.To ensure the accuracy and reproducibility of the BCR::ABL1 p210 test,it is imperative for every laboratory to enhance their daily quality control practices.