Molecular analysis of T cell receptor V beta chain to detect leukemia cell clonality in patients by adaptor ligation-mediated polymerase chain reaction.
We have developed a simple and rapid method to analyze the clonality of leukemia cells. After three rounds of amplification by adaptor-ligation polymerase chain reaction (PCR), the cDNA is cut with AluI, HaeIII, RsaI, and Sau3AI, and analyzed by polyacrylamide gel electrophoresis. The size of the restriction fragments is compared to that of the published restriction fragments size each TCR-beta subfamily V region. The sensitivity of adaptor-ligation PCR restriction enzyme analysis (AL-PCR-REA) was 10(-4) MOLT-4 T-ALL cell population in the normal peripheral blood lymphocytes (PBL). Application of AL-PCR-REA to PBL and bone marrow (BM) cells from eight clinical leukemia samples indicated that a detection sensitivity was rather low, but revealed the clonality of all eight clinical samples. This AL-PCR-REA method can detect clonality without the need for either radioisotopes or sequencing procedures.
Matsutani
T
T
Shionogi Biomedical Laboratories, Shionogi & Co., Ltd., Osaka, Japan.
Oishi
C
C
Kaneshige
T
T
Igimi
H
H
Uchida
K
K
Tsuruta
Y
Y
Yoshioka
T
T
Suzuki
R
R
Sakata
T
T
D001483:Base Sequence; D001854:Bone Marrow Cells; D002999:Clone Cells; D011088:DNA Ligases / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D005796:Genes; D006801:Humans; D007938:Leukemia / Q000235:genetics / Q000473:pathology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D000091402:Prohibitins; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D015183:Restriction Mapping
D017931:DNA Primers; D018076:DNA, Complementary; D004273:DNA, Neoplasm; C000717331:PHB2 protein, human; D000091402:Prohibitins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011088:DNA Ligases