Droplet Digital PCR-Based Chimerism Analysis for Primary Immunodeficiency Diseases
Droplet Digital PCR-Based Chimerism Analysis for Primary Immunodeficiency Diseases
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DOI：
10.1007/s10875-018-0497-8
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发表时间：
2018-04-01
影响因子：
9.1
通讯作者：
Morio, Tomohiro
中科院分区：
文献类型：
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作者：
Okano, Tsubasa;Tsujita, Yuki;Morio, Tomohiro
为准确评价造血干细胞移植（HSCT）患者供/受者或男女嵌合状态，设计了SRY和RPP 30的液滴数字聚合酶链反应（ddPCR）检测HSCT患者供/受者或男女嵌合状态。我们还开发了突变特异性ddPCR的四个主要免疫缺陷diseases.The男性/女性嵌合体分析使用ddPCR的准确性进行了确认，通过比较结果与传统的方法（荧光原位杂交和短串联重复序列PCR）和评估稀释试验。特别是，我们发现这种方法对于分析小样品是有用的。因此，该方法可用于患者样本，特别是在移植后早期分选的白细胞亚群。4种突变特异性ddPCR均能准确检测移植后嵌合体，基于ddPCR的男性/女性嵌合体分析和突变特异性ddPCR均适用于所有HSCT，这些简单的方法有助于追踪移植后嵌合体，尤其是疾病特异性小白细胞组分。
In the current study, we aimed to accurately evaluate donor/recipient or male/female chimerism in samples from patients who underwent hematopoietic stem cell transplantation (HSCT).We designed the droplet digital polymerase chain reaction (ddPCR) for SRY and RPP30 to detect the male/female chimerism. We also developed mutation-specific ddPCR for four primary immunodeficiency diseases.The accuracy of the male/female chimerism analysis using ddPCR was confirmed by comparing the results with those of conventional methods (fluorescence in situ hybridization and short tandem repeat-PCR) and evaluating dilution assays. In particular, we found that this method was useful for analyzing small samples. Thus, this method could be used with patient samples, especially to sorted leukocyte subpopulations, during the early post-transplant period. Four mutation-specific ddPCR accurately detected post-transplant chimerism.ddPCR-based male/female chimerism analysis and mutation-specific ddPCR were useful for all HSCT, and these simple methods contribute to following the post-transplant chimerism, especially in disease-specific small leukocyte fractions.