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Real‐time PCR quantification of haematopoietic chimerism after transplantation: a comparison between TaqMan and hybridization probes technologies
Author(s) -
MARTINEZLOPEZ J.,
CROOKE A.,
GRANDE S.,
AYALA R.,
JIMÉNEZVELASCO A.,
GAMARRA S.,
MENEU J. C.,
GILSANZ F.
Publication year - 2010
Publication title -
international journal of laboratory hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.705
H-Index - 55
eISSN - 1751-553X
pISSN - 1751-5521
DOI - 10.1111/j.1751-553x.2009.01166.x
Subject(s) - taqman , transplantation , real time polymerase chain reaction , haematopoiesis , hematopoietic stem cell transplantation , microbiology and biotechnology , confidence interval , polymerase chain reaction , coefficient of variation , biology , medicine , stem cell , genetics , chemistry , chromatography , gene
Summary 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.