Abstract
Background: Analysis of the relative amounts of donor and recipient DNA in bone marrow after bone marrow transplantation is frequently used to determine the statusof the transplant. We studied the performance of an assay to quantify chimerism based on amplification of the D1S80 variable number tandem repeat marker by PCR with detection of PCR products by capillary electrophoresis (CE).
Methods and Results: Samples from potential bone marrow donors and recipients were analyzed separately and in mixtures to simulate various degrees of chimerism from 10% to 90% and subjected to PCR/CE analysis. There was excellent agreement between the measured and known relative proportions of DNA components in chimeric samples. The lower limit of sensitivity for detection of chimerism was 1%; between-runs coefficients of variation were <5%.
Conclusions: Amplification of the D1S80 minisatellite by PCR with CE detection is a reliable method for determination of the relative contribution of different DNAs in mixed samples. This method is fast, quantitative, and extremely reproducible.
Similar content being viewed by others
References
Kloosterman AD, Budowle B, Daselaar P: PCR-amplification and detection of the human D1S80 VNTR locus. Int J Legal Med 1993;105: 257–264
Lorente M, Lorente JA, Wilson MR, Budowle B, Villanueva E: Spanish population data on seven loci:D1S80, D17S5, HUMTH01, HUMVWA, ACTBP2, DS1S11 and HLA-DQA1. Forensic Sci Int 1997;86: 163–171
Budowle B, Chakraborty R, Giusti AM, Eisenbert AJ, Allen RC: Analysis of the VNTR D1S80 by the PCR followed by high-resolution PAGE. Am J Hum Genet 1991;48: 137–144
Budowle B, Baecheel FS, Smerick JB, et al: D1S80 population data in African Americans, Caucasians, Southeastern Hispanics, Southwestern Hispanics, and Orientals. J Forensic Sci 1995;40: 38–44
Zhang N, Yeung ES: Genetic typing by capillary electrophoresis with the allelic ladder as an absolute standard. Anal Chem 1996;68: 2927–2931
Vainer M, Enad S, Dolnik V, et al.: Short tandem repeat typing by capillary array electrophoresis: Comparison of sizing accuracy and precision using different buffer systems. Genomics 1997;41: 1–9
Isenberg AR, McCord BR, Koons BW, Budowle B, Allen RO: DNA typing of a polymerase chain reaction amplified D1S80/amelogenin multiplex using capillary electrophoresis and a mixed entangled polymer matrix. Electrophoresis 1996;17: 1505–1511
Gelfi G, Orsi A, Righetti PG, Brancolini V, Cremonesi L, Ferrari M: Capillary zone electrophoresis of polymerase chain reaction-amplified DNA fragments in polymer networks: The case of GATT microsatellites in cystic fibrosis. Electrophoresis 1994; 15: 641–643
Nesi M, Righetti PG, Patrosso MC, Ferlini A, Chiari M: Capillary electrophoresis of polymerase chain reaction-amplified products in polymer networks: The case of Kennedy’s disease. Electrophoresis 1994;15: 644–646
Baba Y, Tomisake R, Sumita C, et al.: Rapid typing of variable number of tandem repeat locus in the human apolipoprotein B gene for DNA diagnosis of heart disease by polymerase chain reaction and capillary electrophoresis. Electrophoresis 1995;16: 1437–1440
Morrison C, Gannon F: The impact of the PCR plateau phase on quantitative PCR. Biochim Biophys Acta 1994;1219:493–498
Zhou M, Sheldon S, Akel N, Killeen AA: Chromosomal aneuploidy in leukemic blast crisis: A potential source of error in interpretation of bone marrow engraftment analysis by VNTR amplification. Mol Diag 1999;4: 153–157
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Jone, C.M., Akel, N. & Killeen, A.A. Evaluation of Chimerism in DNA Samples by PCR Amplification of D1S80 With Detection by Capillary Electrophoresis. Molecular Diagnosis 5, 101–105 (2000). https://doi.org/10.1007/BF03262028
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF03262028