4.2 Article

Ultra-Sensitive Droplet Digital PCR for the Assessment of Microchimerism in Cellular Therapies

期刊

BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION
卷 24, 期 5, 页码 1069-1078

出版社

ELSEVIER SCIENCE INC
DOI: 10.1016/j.bbmt.2017.12.802

关键词

Chimerism; Microtransplantation; Adoptive T-cell therapy; Virus-specific T cells; ddPCR

资金

  1. Australian National Health and Medical Research Council (NHMRC) [PG1061252, PG1032431]
  2. NSW Health Pathology (Institute of Clinical Pathology and Medical Research)
  3. Leukaemia Foundation of Australia
  4. Leukamia and Lymphoma Society
  5. Westmead Charitable Trust

向作者/读者索取更多资源

Current techniques to assess chimerism after hematopoietic stem cell transplantation (HSCT) are limited in both sensitivity and precision. These drawbacks are problematic in the context of cellular therapies that frequently result in microchimerism (donor chimerism <1%). We have developed a highly sensitive droplet digital PCR (ddPCR) assay using commercially available regents with good performance throughout the range of clinically relevant chimerism measurements, including microchimerism. We tested the assay using spiked samples of known donor-recipient ratios and in clinical samples from HSCT recipients and patients enrolled on clinical trials of microtransplantation and third-party virus-specific T cells (VSTs). The levels of detection and quantification of the assay were.008% and.023%, with high levels of precision with samples of DNA content ranging from 1 to 300 ng DNA. From the panel of 29 insertion-deletion probes multiple informative markers were found for each of 43 HSCT donor-recipient pairs. In the case of third-party cellular therapies in which there were 3 DNA contributors (recipient, HSCT donor, and T-cell donor), a marker to detect the cellular product in a background of recipient and donor cells was available for 11 of 12 cases (92%). Chimerism by ddPCR was able to quantify chimerism in HSCT recipients and comparison against standard STR analysis in 8 HSCT patients demonstrated similar results, with the advantage of fast turnaround time. Persistence of donor microchimerism in patients undergoing microtransplantation for acute myeloid leukemia was detectable for up to 57 days in peripheral blood and bone marrow. The presence of microtransplant product DNA in bone marrow T cells after cell sorting was seen in the 1 patient tested. In patients receiving third-party VSTs for treatment of refractory viral infections, VST donor DNA was detected at low levels in 7 of 9 cases. ddPCR offers advantages over currently available methods for assessment of chimerism in standard HSCT and cellular therapies. (C) 2018 American Society for Blood and Marrow Transplantation.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.2
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据