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Clonal analysis of hematopoietic cells using a novel polymorphic site of the X chromosome
Author(s) -
Okamoto Takahiro,
Okada Masaya,
Wada Hiroshi,
Kanamaru Akihisa,
Kakishita Eizo,
Hashimoto Tomoko,
Furuyama Junichi
Publication year - 1998
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/(sici)1096-8652(199808)58:4<263::aid-ajh2>3.0.co;2-o
Subject(s) - loss of heterozygosity , biology , xq28 , primer (cosmetics) , x chromosome , population , polymerase chain reaction , microbiology and biotechnology , haematopoiesis , myeloid , x inactivation , microsatellite , genetics , gene , immunology , stem cell , allele , medicine , chemistry , organic chemistry , environmental health
Clonality of hematopoietic cells on a smale scale (nanogram amounts of DNA) can be detected by X‐chromosome inactivation using the polymerase chain reaction (PCR). The human androgen‐receptor gene (HUMARA) has a polymorphic short tandem repeat (STR), and has generally been used for clonality analysis since heterozygosity for the gene occurs in 90% of caucasian females. We examined heterozygosity of the STR on HUMARA in 110 Japanese females and found heterozygosity in 74 of 110 (67%). To examine for hematologic clonality in females with HUMARA homozygosity, we used a primer specific for a novel polymorphic STR site between DXS15 and DXS134 (DXS15‐134) on Xq28. Heterozygosity for this site was found in 50 of 110 females (46%). Clonality of the hematopoietic cells was detected in 91 of 110 females (83%) using PCR of either the STR sites on HUMARA or DXS15‐134. The X‐inactivation patterns using PCR of DXS15‐134 corresponded exactly with those obtained using PCR of HUMARA in 18 females who were heterozygous for both DXS15‐134 and HUMARA. Using PCR of DXS15‐134, we examined the clonality of bone marrow cells separated by flow cytometry in a patient with erythroleukemia (M6). Clonality was found not only in myeloid lineage cells but also in B lymphocytes. The clonality assay for DXS15‐134 may be useful to assess for clonality of hematopoietic cells in the Japanese population, when combined with the HUMARA assay. Am. J. Hematol. 58:263–266, 1998. © 1998 Wiley‐Liss, Inc.