Detection of T-cell receptor delta gene rearrangement by clonal specific polymerase chain reaction.
A sensitive and specific technique to detect minimal residual disease for T-cell malignancies was explored. Southern analysis and polymerase chain reaction (PCR) were used to detect the rearranged V-D-J segment of T-cell receptor delta (TCR delta) gene from malignant cell specimens of patients with leukemia and lymphoma of T-cell lineage. The PCR product was sequenced and from the DNA sequences of the V-D-J region, a 3' antisense primer was designed and synthesized for clonal specific PCR (CS-PCR). Seven of the 22 T-ALL (32%) and 5 of 18 (28%) T-cell lymphoma showed clonal rearrangement by Southern analysis. Six of the 7 (86%) T-ALL and 4 of the 5 (80%) T-cell lymphoma which were Southern positive were also positive by TCR delta PCR. The PCR products of four cases of T-ALL showing clonal pattern by TCR delta PCR amplification were successfully sequenced and CS-PCR amplification performed. CS-PCR detected with confidence specific clonal rearrangement in a mixture containing 0.003% of malignant cells. Marrow specimens obtained at diagnosis and subsequent follow-ups from the 4 T-ALL patients were studied by Southern analysis, TCR delta PCR and CS-PCR. The first patient was in continuous morphological complete remission for more than 3 years and had persistently negative Southern, TCR delta PCR and CS-PCR results on follow-up. Initial follow-up marrow samples from the second patient had persistently positive CS-PCR results while they were still morphologically and TCR delta PCR negative and the patient had a frank leukemic relapse at 18 months. The other two patients had persistent disease by conventional morphological examination, Southern analysis, TCR delta PCR and CS-PCR studies were all positive as expected. CS-PCR is a highly specific and sensitive technique in detecting minimal residual disease for T-cell malignancies. Its potential applications warrant further clinical evaluation and correlation.
Chan
D W
DW
Dept. of Medicine, Queen Mary Hospital, Pokfulam, H.K.
Liang
R
R
Chan
V
V
Kwong
Y L
YL
Chan
T K
TK
D001483:Base Sequence; D001853:Bone Marrow / Q000473:pathology; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D005500:Follow-Up Studies; D016694:Gene Rearrangement, delta-Chain T-Cell Antigen Receptor; D006801:Humans; D015458:Leukemia, T-Cell / Q000235:genetics* / Q000276:immunology / Q000401:mortality / Q000473:pathology; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000235:genetics / Q000276:immunology; D016399:Lymphoma, T-Cell / Q000235:genetics* / Q000276:immunology / Q000401:mortality / Q000473:pathology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011379:Prognosis; D012680:Sensitivity and Specificity; D015996:Survival Rate
D017931:DNA Primers