Quantitative analysis of chimerism after allogeneic hematopoietic stem cell transplantation with molecular genetic methods

Onkogematologiâ

Ye. V. Volochnik¹  O. V. Aleynikova¹  Yu. E. Mareiko¹  T. V. Savitskaya¹  V. A. Lavrinenko¹ 

[1] Republican Scientific Center of Pediatric Oncology, Hematology and Immunology, Ministry of Health of Republic of Belarus, Minsk;

关键词: chimerism;    short tandem repeats;    insertion / deletion polymorphisms;    polymerase chain reaction;    fluorescence in situ hybridization;    hematopoietic stem cell transplantation;   

DOI  :  10.17650/1818-8346-2014-9-2-29-36

来源: DOAJ

【 摘 要 】

Quantitative monitoring of chimerism after allogeneic hematopoietic stem cell transplantation (HSCT) by molecular methods has becomea significant diagnostic tool in detection of engraftment / graft failure, predicting rejection and disease relapse. Despite the great utility of chimerism analysis there is not a unique standard method for its quantification. The objective of the present investigation was to compare perspective methods multiplex short tandem repeat polymerase chain reaction (STR-PCR) and real-time PCR insertion / deletion polymorphisms (InDel-PCR) for the quantification of chimerism after HSCT. We performed a study analyzing the chimerism status in 60 patients by STR-PCR and by InDel-PCR. Recipient / donor discrimination was possible with STR-PCR in all patient-donor pairs (100 %), whereas informative alleles for recipient were found in 88 % pairs with InDel-PCR. The sensitivity (detection limit) of STR-PCR and InDel-PCR was 1–5 % and more than 0.01 % donor cells correspondingly. The accuracy of quantification was higher for STR-PCR than for InDel-PCR, when level of donor chimerism was 3–97 %. These methods can be successfully used to determine chimerism after allogeneic HSCT. Considering the higher sensitivity and quantification accuracy of InDel-PCR it should be chosen if donor chimerism level less 5 % or more 95 % and in other cases STR-PCR should be chosen.

【 授权许可】

Unknown