Deteksi ekspresi Gen Fusi bcr-abl dan pml-rara :: Pada kasus leukemia dewasa di RS Dr. Sardjito

HERNINGTYAS, Elizabeth Henny, Prof.Dr.dr. Sofia Mubarika, MMedSc.,Ph.D

2004 | Tesis | S2 Bioteknologi

Leukemia merupakan keganasan darah karena proliferasi berlebihan dari sel hematopoetik atau limfoid yang dapat mengenai semua orang, jenis kelamin dan kelompok umur. Kasus leukemia di Indonesia cukup banyak (± 7000 kasus/tahun) dengan angka kematian 83,6%. Namun, diagnosis yang dilakukan masih berdasarkan pada klasifikasi FAB, padahal berdasarkan klasifikasi WHO (2000), deteksi secara sitogenetik dan biologi molekular sangat berperan dalam penentuan diagnosis klonal penyakit dan pemilihan terapi yang tepat sehingga perlu dikembangkan deteksi secara molekular di Indonesia khususnya di RS Dr. Sardjito. Tujuan penelitian ini untuk mendapatkan profil ekspresi fusi gen bcr-abl dan pml-rara pada penderita leukemia dewasa di RS dr. Sardjito. Kriteria inklusi: penderita dewasa yang didiagnosis leukemia (CML, ALL, AML M3) oleh dokter di RS berdasarkan pemeriksaan klinis, laboratoris, morfologi darah tepi, punksi sumsum tulang, sitokimia maupun imunofenotiping. Darah penderita diambil sel mononuklearnya dengan pemisahan secara gradien sentrifugasi dengan Ficoll-hypaque. Isolasi RNA dilakukan dengan Trizol dan diubah menjadi cDNA dengan RT-reaction. Gen β-actin digunakan sebagai kontrol internal, dan gen bcr-abl maupun pml-rara diamplifikasi secara nested PCR . Dari 4 kasus yang diperoleh yaitu CML (2 kasus), ALL (1 kasus) dan AML M3 (1 kasus) ternyata satu kasus CML pasca terapi memberikan hasil positif pada gen fusi bcr-abl.

Leukemia is hematologic malignancy caused by excess proliferation of hematopoetic or lymphoid cells that affect everybody, ages and sex. Leukemia cases in Indonesia is a lot enough (± 7000 cases/year) with mortality rate 83,6%. Diagnosis is based on FAB classification morphology. The fact, WHO classification 2000 has been used worldwide based on cytogenetic and molecular biology profile can define the clonal diseases more precisely and choose the adequate therapy. The aim of the research was to get the expression profiles of bcr-abl and pmlrara fusion genes in adult leukemia at Sardjito Hospital. The inclusion criterias were adult patient suspected or had been diagnosed as leukemia (CML, ALL, AML M3) by hospital doctors based on the clinical examination, laboratory tests, peripheral blood morphology, bone marrow punction, cytochemistry or imunophenotyping. Mononuclear cell of the samples were separated by Ficoll-hypaque gradient centrifugation. RNA was isolated by Trizol and converted to cDNA by RT reaction. β-actin gene was used as internal control and bcr-abl and pml-rara gene were amplified by nested PCR. From 4 cases used in this study namely CML (2 cases), ALL (1case) and AML M3 (1 case) showed that one post therapy CML case gave positive result for bcr-abl fusion gene.

Kata Kunci : Bioteknologi,Deteksi Molekular Leukemia,Gen bcr,abl,adult leukemia, fusion gene, bcr-abl, pml-rara, WHO classification