Evaluation of 16 SNPs allele-specific to quantify post hSCT chimerism by SYBR green-based qRT-PCR

J Clin Pathol. 2013 Mar;66(3):238-42. doi: 10.1136/jclinpath-2012-201224. Epub 2013 Jan 2.

Abstract

The importance of monitoring post haematopoietic stem cell transplantation (hSCT) chimerism has been defined in numerous publications. Single-nucleotide polymorphisms (SNPs) are molecular markers that vary significantly among different populations. Allied to a very sensible technique, SNP assays seem to be very sensitive (0.001%) when post hSCT chimerism is measured. However, well known SNP frequencies are limited to certain populations, mainly in countries where there is a high level of diversity in its population, therefore restricting their use worldwide. Amplification by SYBR green based quantitative real time PCR of eight pairs of allele-specific SNPs (MLH-1, PECAM-1, ICAM-1, SUR-1, HA-1, rs715405, rs713503, rs2296600) was conducted in 88 patient/donor pairs, who underwent allogeneic myeloablative or non-myeloablative hSCT. One informative allele was detected in at least 42% (n=37) of the samples; 20% (n=18) had at least two informative alleles; 10% (n=9) had at least three informative alleles; 9% (n=8) had more than three informative alleles and 18% (n=16) showed no informative allele at all. Overall, the frequency of informative alleles for these SNPs in the Brazilian population was very low. Consequently, the amount of information attained reached 9% of those expected, being able to discriminate only eight pairs of donor/recipient samples with more than three informative alleles, making them useless for the quantification of chimerism in our routine.

MeSH terms

  • Adult
  • Benzothiazoles
  • DNA / chemistry
  • Diamines
  • Female
  • Fluorescent Dyes
  • Genetic Markers / genetics*
  • Genotype
  • Hematopoiesis / genetics
  • Hematopoietic Stem Cell Transplantation*
  • Humans
  • Living Donors
  • Male
  • Oligonucleotide Array Sequence Analysis / methods*
  • Organic Chemicals / chemistry
  • Polymorphism, Single Nucleotide*
  • Quinolines
  • Real-Time Polymerase Chain Reaction / methods*
  • Transplantation Chimera / genetics*
  • Transplantation, Homologous
  • Unrelated Donors

Substances

  • Benzothiazoles
  • Diamines
  • Fluorescent Dyes
  • Genetic Markers
  • Organic Chemicals
  • Quinolines
  • SYBR Green I
  • DNA