Multiprimer--PCR for screening of IgH and T-cell receptor rearranged genes in acute lymphoblastic leukemia.

OBJECTIVE

To develop a multiprimer-polymerase chain reaction (multiprimer-PCR) method for detecting immunoglobulin heavy chain (IgH) and T-cell receptor (TCR) rearranged genes in the same amplification.

METHODS

Multiprimer-PCR protocol, a mixture of four different primers used in the same reaction detected IgH CDR-III and TCR V gamma I-J gamma Ligenetic rearrangements in 40 acute lymphoblastic leukemia (ALL) patients.

RESULTS

Thirteen cases were found to have IgH CDR-III and TCR V gamma I-J gamma genetic rearrangements. Ten cases were found to have TCR V gamma I-J gamma genetic rearrangements. Thirteen cases were found to have IgH CDR-III genetic rearrangements. The amplification result of multiprimer-PCR was the same as separate amplification. The sensitivity of multiprimer-PCR was 10(-4)-10(-5) DNA level, which was the same as separate PCR amplification. All positive cases with IgH rearranged gene were confirmed by Southern blot.

CONCLUSION

Multiprimer-PCR could screen two distinct rearranged genes in a single amplification, and is more suitable for screening of specimens collected at diagnosis.