Department of Laboratory Medicine, Shinshu University Hospital, Asahi, Matsumoto, Japan.
Clin Chim Acta. 2012 Feb 18;413(3-4):516-9. doi: 10.1016/j.cca.2011.11.017. Epub 2011 Nov 25.
Quantitative evaluation of minimal residual disease (MRD) following hematopoietic stem cell transplantation (HSCT) is indispensable for patients with hematological malignancies. In addition to established MRD markers such as immunoglobulin and T-cell receptor gene rearrangements, fusion genes, or aberrantly expressed genes, single nucleotide mutations are considered one of the MRD markers that reflect the malignant cell clone.
We compared the quantity of mutant genes by allele-specific quantitative polymerase chain reaction (AS-qPCR) for single nucleotide mutations (TP53 410T>A and PTPN11 1508G>A) with the percentage of autologous DNA by short tandem repeat (STR)-PCR.
Following HSCT, the quantity of mutant genes detected by AS-qPCR correlated with the percentage of autologous DNA assessed by the STR-PCR. Moreover, mutant DNAs were detected at a quantifiable level before relapse, whereas the percentage of autologous DNA was less than 5%, that is, complete chimerism.
The AS-qPCR approach for single nucleotide mutations was accurate and highly sensitive for monitoring pre-transplantation as well as post-transplantation MRD. AS-qPCR for single nucleotide mutation is suitable for monitoring MRD in patients who lack previously established MRD markers.
对于血液系统恶性肿瘤患者，造血干细胞移植（HSCT）后微小残留病（MRD）的定量评估是不可或缺的。除了已确立的 MRD 标志物，如免疫球蛋白和 T 细胞受体基因重排、融合基因或异常表达的基因外，单核苷酸突变被认为是反映恶性细胞克隆的 MRD 标志物之一。
我们通过等位基因特异性定量聚合酶链反应（AS-qPCR）比较了单核苷酸突变（TP53 410T>A 和 PTPN11 1508G>A）的突变基因数量与短串联重复（STR）-PCR 评估的自体 DNA 百分比。
HSCT 后，AS-qPCR 检测到的突变基因数量与 STR-PCR 评估的自体 DNA 百分比相关。此外，在复发前可检测到可定量的突变 DNA，而自体 DNA 百分比小于 5%，即完全嵌合体。
用于单核苷酸突变的 AS-qPCR 方法对于监测移植前和移植后 MRD 是准确和高度敏感的。用于单核苷酸突变的 AS-qPCR 适用于缺乏先前建立的 MRD 标志物的患者的 MRD 监测。
