Digital PCR to assess hematopoietic chimerism after allogeneic stem cell transplantation

Experimental Hematology
Tanja StahlB Fehse

Abstract

Analysis of hematopoietic chimerism after allogeneic stem cell transplantation represents a crucial method to evaluate donor-cell engraftment. Whereas sensitivity of classical approaches for chimerism monitoring is limited to ≥1%, quantitative polymerase chain reaction (qPCR)-based techniques readily detect one patient cell in >1,000 donor cells, thus facilitating application of chimerism assessment as a surrogate for minimal residual disease. However, due to methodologic specificities, qPCR combines its high sensitivity with limited resolution power in the state of mixed chimerism (e.g., >10% patient cells). Our aim was to overcome this limitation by employing a further development of qPCR, namely digital PCR (dPCR), for chimerism analysis. For proof-of-principle, we established more than 10 dPCR assays detecting Indel polymorphisms or Y-chromosome sequences and tested them on artificial cell mixtures and patient samples. Employing artificial cell mixtures, we found that dPCR allows exact quantification of chimerism over several orders of magnitude. Digital PCR results proved to be highly reproducible (deviation <5%), particularly in the "difficult" range of mixed chimerism. Excellent performance of the new assays was confirme...Continue Reading

References

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Citations

Sep 13, 2016·International Journal of Molecular Sciences·Tanja StahlBoris Fehse
Jan 23, 2017·Molecular Therapy : the Journal of the American Society of Gene Therapy·Malte MohmeBoris Fehse
Jun 22, 2019·International Journal of Laboratory Hematology·Thomas MikaSusanne Klein-Scory
Feb 25, 2019·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·Junpeng ZhaoLunan Wang
Jul 10, 2019·Stem Cells International·Catherine A LombardEtienne M Sokal
Jan 11, 2019·International Journal of Molecular Sciences·Christina RautenbergThomas Schroeder
May 4, 2018·Bone Marrow Transplantation·Lea SellmannLothar Hambach
Aug 27, 2019·Frontiers in Oncology·Irene Della StarzaRobin Foà
May 16, 2019·Bone Marrow Transplantation·Juergen FoellSelim Corbacioglu
May 6, 2020·International Journal of Molecular Sciences·Nicoletta CoccaroFrancesco Albano
Oct 22, 2020·Bone Marrow Transplantation·Tongted P DasAndrew Spencer
Feb 15, 2020·Molecular Therapy. Methods & Clinical Development·Boris FehseFrancis A Ayuk
Nov 18, 2021·Molecular Therapy. Methods & Clinical Development·Anita BadbaranBoris Fehse

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