title: A novel clonality assay based on transcriptional analysis of the active X chromosome.
abstract: The clonal origin of malignancy and hematopoiesis is a principal tenet of modern biology and medicine. This paper describes a highly specific and sensitive assay for the detection of clonality in cells and cell lineages suitable for studies in a large proportion of females. The specific ligase chain and/or ligase detection reactions (LCR/LDR) are utilized at a polymorphic glucose-6-phosphate dehydrogenase (G-6-PD) locus for discrimination of the mRNA transcripts of the active X chromosome. This combination approach circumvents problems encountered with other currently used assays of clonality based either on peptide G-6-PD polymorphism or on DNA methylation differences between the active and inactive X chromosomes. The veracity of this assay was verified by analysis of 19 random healthy females as well as by the study of hemopoietic and nonhemopoietic tissues from a patient with clonal hemopoiesis/polycythemia vera. Furthermore, we demonstrate that the G-6-PD locus used in our clonal assay does not display marked differences in methylation between the active and the inactive X chromosomes.
authors.affiliation: Division of Hematology/Oncology, University of Alabama, Birmingham 35294.
authors.forename: J T
authors.initials: JT
authors.lastname: Prchal
authors.forename: Y L
authors.initials: YL
authors.lastname: Guan
mesh_terms: D000483:Alleles; D002999:Clone Cells; D004273:DNA, Neoplasm / Q000378:metabolism; D005260:Female; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D006410:Hematopoiesis; D006801:Humans; D008745:Methylation; D014411:Neoplastic Stem Cells; D011087:Polycythemia Vera / Q000201:enzymology / Q000235:genetics* / Q000473:pathology*; D014158:Transcription, Genetic; D014960:X Chromosome
chemical_list: D004273:DNA, Neoplasm; D005954:Glucosephosphate Dehydrogenase