Real-time PCR quantification of haematopoietic chimerism after transplantation: a comparison between TaqMan and hybridization probes technologies

Int J Lab Hematol. 2010 Feb;32(1 Pt 1):e17-25. doi: 10.1111/j.1751-553X.2009.01166.x. Epub 2009 May 12.

Abstract

This study aimed to compare the sensitivity and accuracy of two methods of quantitative real-time polymerase chain reaction (qrt-PCR), in order to determine haematopoietic chimerism (CH): single nucleotide polymorphisms using TaqMan (TM) probes and insertion/deletion polymorphisms using Hybridization (Hyb) probes. A total of 106 samples from 20 patients who underwent allogenic stem cell transplantation (n = 14) or live-donor liver transplantation (n = 6) were studied. The mean level of chimerism was 8.37% for the TM method and 7.73% in the Hyb method, which was not significantly different (P = 0.69). The Pearson correlation coefficient between the two methods was r = 0.91 (P < 0.001). The estimation of the regression line, using the Passing and Balbock method was Intercept A -0.0381 [95% confidence interval (CI) -0.1265 to 0.0296) and Slope B: 1.04609(95% CI 0.9349-1.161). Bland-Altman data showed that the standard deviations, which differed between the two methods (%Hyb-%TM), were 0.98 and -1.28. The accuracy and sensitivity of qrt-PCR chimerism is independent of the method used if the optimization is adequate and satisfies the criteria for adequate study. Real-time PCR, independent of the method adopted, is a very good tool for study levels of CH.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / genetics
  • Gene Deletion
  • Genetic Markers / immunology
  • Hematopoietic Stem Cell Transplantation
  • Humans
  • Liver Transplantation / immunology*
  • Mutagenesis, Insertional
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single Nucleotide
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Transplantation Chimera / immunology*
  • Transplantation, Homologous / immunology*

Substances

  • Genetic Markers
  • DNA