title: Real-time PCR quantification of haematopoietic chimerism after transplantation: a comparison between TaqMan and hybridization probes technologies.
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.
authors.affiliation: Haematology Service, Hospital 12 de Octubre, Madrid, Spain. jmartinezlo@yahoo.es
authors.forename: J
authors.initials: J
authors.lastname: Martínez-López
authors.forename: A
authors.initials: A
authors.lastname: Crooke
authors.forename: S
authors.initials: S
authors.lastname: Grande
authors.forename: R
authors.initials: R
authors.lastname: Ayala
authors.forename: A
authors.initials: A
authors.lastname: Jiménez-Velasco
authors.forename: S
authors.initials: S
authors.lastname: Gamarra
authors.forename: J C
authors.initials: JC
authors.lastname: Meneu
authors.forename: F
authors.initials: F
authors.lastname: Gilsanz
mesh_terms: D004247:DNA / Q000235:genetics; D017353:Gene Deletion; D005819:Genetic Markers / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D016031:Liver Transplantation / Q000276:immunology*; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018183:Transplantation Chimera / Q000276:immunology*; D014184:Transplantation, Homologous / Q000276:immunology*
chemical_list: D005819:Genetic Markers; D004247:DNA