Rapid single nucleotide polymorphism based method for hematopoietic chimerism analysis and monitoring using high-speed droplet allele-specific PCR and allele-specific quantitative PCR

Clinica Chimica Acta; International Journal of Clinical Chemistry
Chiaki TairaTakayuki Honda

Abstract

Chimerism analysis is important for the evaluation of engraftment and predicting relapse following hematopoietic stem cell transplantation (HSCT). We developed a chimerism analysis for single nucleotide polymorphisms (SNPs), including rapid screening of the discriminable donor/recipient alleles using droplet allele-specific PCR (droplet-AS-PCR) pre-HSCT and quantitation of recipient DNA using AS-quantitative PCR (AS-qPCR) following HSCT. SNP genotyping of 20 donor/recipient pairs via droplet-AS-PCR and the evaluation of the informativity of 5 SNP markers for chimerism analysis were performed. Samples from six follow-up patients were analyzed to assess the chimerism via AS-qPCR. These results were compared with that determined by short tandem repeat PCR (STR-PCR). Droplet-AS-PCR could determine genotypes within 8min. The total informativity using all 5 loci was 95% (19/20). AS-qPCR provided the percentage of recipient DNA in all 6 follow-up patients without influence of the stutter peak or the amplification efficacy, which affected the STR-PCR results. The droplet-AS-PCR had an advantage over STR-PCR in terms of rapidity and simplicity for screening before HSCT. Furthermore, AS-qPCR had better accuracy than STR-PCR for quantific...Continue Reading

References

Aug 1, 1992·PCR Methods and Applications·R S ChaW G Thilly
Jul 29, 2008·Bone Marrow Transplantation·J L Liesveld, P G Rothberg
Dec 6, 2008·The Journal of Molecular Diagnostics : JMD·Egle GineikieneLaimonas Griskevicius
Dec 28, 2010·Clinica Chimica Acta; International Journal of Clinical Chemistry·Ding-Ping ChenChien-Feng Sun
Dec 14, 2011·Clinica Chimica Acta; International Journal of Clinical Chemistry·A OdriozolaM T Zarrabeitia
Jul 4, 2012·Clinica Chimica Acta; International Journal of Clinical Chemistry·Kazuyuki MatsudaTakayuki Honda
May 21, 2013·Clinica Chimica Acta; International Journal of Clinical Chemistry·Chiaki TairaTakayuki Honda

❮ Previous
Next ❯

Citations

Apr 6, 2020·International Journal of Legal Medicine·Tikumphorn SathirapatyaKornkiat Vongpaisarnsin
Mar 16, 2017·Journal of Clinical Laboratory Analysis·Chiaki TairaTakayuki Honda
Jun 4, 2019·FEMS Microbiology Reviews·UNKNOWN Consortium OPATHY, Toni Gabaldón

❮ Previous
Next ❯

Related Concepts

Related Feeds

Blood And Marrow Transplantation

The use of hematopoietic stem cell transplantation or blood and marrow transplantation (bmt) is on the increase worldwide. BMT is used to replace damaged or destroyed bone marrow with healthy bone marrow stem cells. Here is the latest research on bone and marrow transplantation.

Allogenic & Autologous Therapies

Allogenic therapies are generated in large batches from unrelated donor tissues such as bone marrow. In contrast, autologous therapies are manufactures as a single lot from the patient being treated. Here is the latest research on allogenic and autologous therapies.