A CRISPR-based assay for the detection of opportunistic infections post-transplantation and for the monitoring of transplant rejection
A CRISPR-based assay for the detection of opportunistic infections post-transplantation and for the monitoring of transplant rejection
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DOI：
10.1038/s41551-020-0546-5
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发表时间：
2020-04-13
影响因子：
28.1
通讯作者：
Collins, James J.
中科院分区：
文献类型：
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作者：
Kaminski, Michael M.;Alcantar, Miguel A.;Collins, James J.
基于 CRISPR-Cas13 的快速且廉价的即时检测可准确检测血液和尿液样本中的机会性病毒 DNA 以及尿液样本中肾移植排斥反应的 mRNA 标记。 在器官移植中，感染和排斥是移植物丢失的主要原因。它们通过免疫抑制的净状态联系在一起。为了更早地诊断和治疗这些疾病，并改善患者的长期预后，需要在移植后监测患者的精细策略。在这里，我们展示了一种基于 CRISPR-Cas13 的快速且廉价的检测方法，可以准确地检测来自患者血液和尿液样本中的 BK 多瘤病毒 DNA 和巨细胞病毒 DNA，以及来自经历急性肾移植排斥的患者尿液样本中水平升高的 CXCL9 信使 RNA（移植排斥标志物）。我们采用了横向流动读数的检测方法，通过简单的可视化，可以对常见的机会性病毒感染和移植物排斥进行移植后监测，并且应该有助于移植后护理点监测。
A fast and inexpensive point-of-care assay based on CRISPR-Cas13 accurately detects the DNA of opportunistic viruses in blood and urine samples as well as an mRNA marker of renal transplant rejection in urine samples.In organ transplantation, infection and rejection are major causes of graft loss. They are linked by the net state of immunosuppression. To diagnose and treat these conditions earlier, and to improve long-term patient outcomes, refined strategies for the monitoring of patients after graft transplantation are needed. Here, we show that a fast and inexpensive assay based on CRISPR-Cas13 accurately detects BK polyomavirus DNA and cytomegalovirus DNA from patient-derived blood and urine samples, as well as CXCL9 messenger RNA (a marker of graft rejection) at elevated levels in urine samples from patients experiencing acute kidney transplant rejection. The assay, which we adapted for lateral-flow readout, enables-via simple visualization-the post-transplantation monitoring of common opportunistic viral infections and of graft rejection, and should facilitate point-of-care post-transplantation monitoring.