1] Public Health Sciences Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave N, Seattle, Washington 98109, USA [2].
Nat Commun. 2013;4:2680. doi: 10.1038/ncomms3680.
T and B cell receptor loci undergo combinatorial rearrangement, generating a diverse immune receptor repertoire, which is vital for recognition of potential antigens. Here we use a multiplex PCR with a mixture of primers targeting the rearranged variable and joining segments to capture receptor diversity. Differential hybridization kinetics can introduce significant amplification biases that alter the composition of sequence libraries prepared by multiplex PCR. Using a synthetic immune receptor repertoire, we identify and minimize such biases and computationally remove residual bias after sequencing. We apply this method to a multiplex T cell receptor gamma sequencing assay. To demonstrate accuracy in a biological setting, we apply the method to monitor minimal residual disease in acute lymphoblastic leukaemia patients. A similar methodology can be extended to any adaptive immune locus.
T 细胞和 B 细胞受体基因座经历组合重排，产生多样化的免疫受体库，这对于识别潜在抗原至关重要。在这里，我们使用多重 PCR 技术，用针对重排的可变和连接片段的混合引物来捕获受体多样性。差异杂交动力学会引入显著的扩增偏差，从而改变通过多重 PCR 制备的序列文库的组成。我们使用合成免疫受体库来识别和最小化这种偏差，并在测序后进行计算去除残留的偏差。我们将这种方法应用于多重 T 细胞受体γ测序检测中。为了在生物学背景下证明其准确性，我们将该方法应用于急性淋巴细胞白血病患者的微小残留病监测。类似的方法可以扩展到任何适应性免疫基因座。
