Zbornik - Društvo genetičara Srbije

V-Usm-18 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 201
Subotica, 30. novembar - 4. decembar 2004.
INICIJALNA DIJAGNOZA I PRAÆENJE MINIMALNE REZIDUALNE
BOLESTI KOD DEÈJIH AKUTNIH LEUKEMIJA UZ POMOÆ PCR
METODOLOGIJE
N. Tošiæ 1 , J. Jovanoviæ 1 , B. Petruèev 1 , L. Krivokapiæ-Dokmanoviæ 2 , D. Janiæ 2 i S. Pavloviæ 1
1
Institut za molekularnu genetiku igenetièki inenjering, Beograd
2
Univerzitetska deèja klinika, Beograd
U našem istraivanju koristili smo multipleks RT-PCR tehniku za detekciju èetiri
najèešæe translokacije kod dece obolele od ALL; t(9; 22) BCR-ABL (p190) fuzioni
transkript (prisutan u 15% sluèajeva), t(1; 19) E2A-PBX1 fuzioni transkript (4%), t(12;
21) TEL-AML1 fuzioni transkript (8%) i t(4; 11) MLL-AF4 fuzioni transkript (0%).
Ukoliko nije detektovana nijedna od ovih translokacija MRB je praæena analizom
klon-specifiènih regiona spajanja rearaniranih gena za IgH i T-æelijkog receptora
(TCR). Za detekciju tri najèešæe genetièke abnormalnosti u AML, PML/RAR fuzioni
transkript za t(15;17) (prisutna u 13% suèajeva), AML1/ETO fuzioni transkript za t(8;21)
(0%) i CBFbeta/MYH11 fuzioni transkript za inv(16) (13%), koristili smo nested
RT-PCR esej. AML pacijenti kod kojih nisu detektovane ove translokacije testirane su na
mutacije u Flt3 genu. Mutacije u Flt3 genu su analizirane PCR-RFLP analizom za D835
taèkastu mutaciju (prisutna u 4% pacijenata) i PCR analizom za Flt3/ITD (internal tandem
duplication) (8%). Metodologija bazirana na PCR analizi predstavlja osetljiv test za
dijagnostikovanje akutnih leukemija i obezbeðuje dodatne markere za praæenje MRB.
INITIAL DIAGNOSIS AND FOLLOW-UP OF MINIMAL RESIDUAL DISEASE
IN CHILDHOOD ACUTE LEUKEMIA USING PCR-BASED METHODOLOGY
In our investigation we used mutiplex RT-PCR tehnique for detection of four most common
translocations in children affected with ALL; t(9; 22) BCR-ABL (p190) fusion transcript
(present in 15% of cases), t(1; 19) E2A-PBX1 fusion transcript (4%), t(12; 21)
TEL-AML1 fusion transcript (8%) and t(4; 11) MLL-AF4 fusion transcript (0%). If none
of the most common translocations in ALL had been available for MRD detection, the
analysis of clone-specific junctional regions of rearranged genes for both IgH and the
T-cell receptor (TCR) was used for the follow up of MRD. For identification of 3 most
commone genetic abnormalities in AML, PML/RAR fusion transcript for t(15;17) (present
in 13% of patients), AML1/ETO fusion transcript for t(8;21) (0%) and
CBFbeta/MYH11 fusion transcript for inv(16) (13%), we used nested RT-PCR assay.
AML patients lacking the most common translocations were tested for the mutations of
Flt3 gene. Mutations of Flt3 gene were analyzed by PCR-RFLP for D835 point mutation
(present in 4% of patients) and by PCR method for Flt3/ITD (internal tandem duplication)
(8%). By using PCR analysis we have provided a sensitive test for the diagnosis of
acute leukemias and additional markers for MRD follow up.