Chimerism post hematopoietic stem cells graft: Benefits of the quantitative PCR technique

The study of chimerism with the short tandem repeats (STRs) is an important tool in the follow-up of recipients after bone marrow transplantation (BMT). The STRs are reliable, reproducible and fast, and detect less than 5% of recipient's cells. However, according to the STR profiles of the dono...

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Veröffentlicht in:Genes and immunity 2005-04, Vol.6, p.S71-S71
Hauptverfasser: Mollet, I, Martin, C, Dubois, V, Gebuhrer, L
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Martin, C
Dubois, V
Gebuhrer, L
description The study of chimerism with the short tandem repeats (STRs) is an important tool in the follow-up of recipients after bone marrow transplantation (BMT). The STRs are reliable, reproducible and fast, and detect less than 5% of recipient's cells. However, according to the STR profiles of the donor and the recipient, one can be hampered to detect the apparition of the recipient's cells and thus make an early prediction of relapse. During the PCR-STR, the Taq polymerase can jump repeated sequences and thus generate "stutters" (ghost peaks) of low amplitude, located at the same level as the recipient's peak. Consequently, such a profile can be misinterpreted and generate a false positive or negative results. In such a case, another strategy can be useful: the real time quantitative PCR using Taqman technology with the specific nucleotide polymorphism (SNP). This technology without PCR competition shows better sensitivity (
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The STRs are reliable, reproducible and fast, and detect less than 5% of recipient's cells. However, according to the STR profiles of the donor and the recipient, one can be hampered to detect the apparition of the recipient's cells and thus make an early prediction of relapse. During the PCR-STR, the Taq polymerase can jump repeated sequences and thus generate "stutters" (ghost peaks) of low amplitude, located at the same level as the recipient's peak. Consequently, such a profile can be misinterpreted and generate a false positive or negative results. In such a case, another strategy can be useful: the real time quantitative PCR using Taqman technology with the specific nucleotide polymorphism (SNP). This technology without PCR competition shows better sensitivity (&lt;0.4%) and linearity. In a retrospective study of the cases of two patients with an unfavourable profile with the STRs markers, we compared the results of chimerism determination performed with both STRs and SNPs techniques. For the first patient, the STRs failed to predict the disease relapse after 14 months of graft when at that time, the SNPs were informative. For the second patient, the STRs did not allow us to draw any conclusion at the time when the level of 7% of recipient cells was reached, while the SNPs allowed us to follow the kinetics of the recipient cells and catch their progressive decrease. These results confirm that when trying to predict as soon as possible the relapse of the disease in BMT recipients, it is mandatory to use a second method to confirm chimerism, in order to clear ambiguities in interpretation linked to the possible presence of "stutters" which may confuse the recipient's profile during PCR-STR.</description><identifier>ISSN: 1466-4879</identifier><language>eng</language><ispartof>Genes and immunity, 2005-04, Vol.6, p.S71-S71</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780</link.rule.ids></links><search><creatorcontrib>Mollet, I</creatorcontrib><creatorcontrib>Martin, C</creatorcontrib><creatorcontrib>Dubois, V</creatorcontrib><creatorcontrib>Gebuhrer, L</creatorcontrib><title>Chimerism post hematopoietic stem cells graft: Benefits of the quantitative PCR technique</title><title>Genes and immunity</title><description>The study of chimerism with the short tandem repeats (STRs) is an important tool in the follow-up of recipients after bone marrow transplantation (BMT). 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title Chimerism post hematopoietic stem cells graft: Benefits of the quantitative PCR technique
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