Streamlined and quantitative detection of chimerism using digital PCR.
Streamlined and quantitative detection of chimerism using digital PCR.
复制标题
利用数字PCR简化和定量检测嵌合现象。
DOI：
10.1038/s41598-022-14467-5
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发表时间：
2022-06-17
影响因子：
4.6
通讯作者：
Nakauchi, Hiromitsu
中科院分区：
文献类型：
--
作者：
Suchy, Fabian P.;Nishimura, Toshiya;Seki, Shinsuke;Wilkinson, Adam C.;Higuchi, Maimi;Hsu, Ian;Zhang, Jinyu;Bhadury, Joydeep;Nakauchi, Hiromitsu
动物嵌合体广泛用于生物医学发现，从发育生物学到癌症研究。然而，嵌合和镶嵌组织中混合细胞类型的准确定量由于样品制备偏倚、转基因沉默、表型相似性和低通量分析管道而变得复杂。在这里，我们已经开发和特点的液滴数字PCR单核苷酸歧视检测常见的白化病和非白化病小鼠品系之间的嵌合体。此外，我们验证了该测定与来自所有实体器官的粗裂解物相容，大大简化了样品制备。与现有方法相比，该嵌合检测试验具有许多额外的优点，包括其稳健性、最小的技术偏倚以及报告制备样品中细胞总数的能力。此外，这里讨论的概念很容易适用于其他基因组基因座，以准确测量任何组织中的混合细胞群体。
Animal chimeras are widely used for biomedical discoveries, from developmental biology to cancer research. However, the accurate quantitation of mixed cell types in chimeric and mosaic tissues is complicated by sample preparation bias, transgenic silencing, phenotypic similarity, and low-throughput analytical pipelines. Here, we have developed and characterized a droplet digital PCR single-nucleotide discrimination assay to detect chimerism among common albino and non-albino mouse strains. In addition, we validated that this assay is compatible with crude lysate from all solid organs, drastically streamlining sample preparation. This chimerism detection assay has many additional advantages over existing methods including its robust nature, minimal technical bias, and ability to report the total number of cells in a prepared sample. Moreover, the concepts discussed here are readily adapted to other genomic loci to accurately measure mixed cell populations in any tissue.
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影响因子：
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通讯作者：
Li S
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通讯作者：
Vandesompele, Jo