DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT
Abstract
Background. The mDx HemaVision kit is a qualitative multiplex and nested Reverse Transcription-Polymerase Chain Reaction (RT-PCR) test designed to detect 28 different translocations or chromosomal rearrangement, found to be specific for particular subtypes of leukemia. The presence or absence of the specific mRNA transcripts or cDNA segments after the reverse transcription of the fusion or abnormal genes, appeared after chromosomal rearrangements, could be determined by the kit.
Patients and methods. The usefulness of the kit was tested on the 26 RNA samples of patients with acute leukemia and seven patients with chronic mieloproliferative diseases and by comparison of the results between mDx HemaVision kit and standardized RT-PCR protocol for the specific mRNA transcripts of the t(9;22)(q34;q11), t(8;21)(q22;q22), t(15;17)(q21;q21) and t(4;11)(q21;q23). The RNA samples were isolated from mononuclear cells of the bone marrow after Ficoll-Paque density gradient centrifugation and with a High Pure RNA isolation kit. The cDNA synthesis and Polymerase Chain Reaction were performed as described in mDx HemaVisoin’s or standardized RT-PCR’s protocols. The PCR products were analyzed by agarose gel electrophoresis, by staining with etidium bromide and by visualization under UV light.
Results. We obtained 100% concordance of the results by both methods. Specific BCR-ABL mRNA transcripts were found in four chronic myeloid leukemia patients, one in B acute lymphoblastic leukemia (B-ALL) and one with bifenotipic leukemia (BAL) patient. AML1-ETO mRNA transcript of the t(8;21)(q22;q22) was identified in two patients with acute myeloid leukemia (AML). The CBF β /MYH11 mRNA transcript specific for inv16(p13;q22) was obtained in AML patient with abnormal eozinofiles in bone marrow. MLL/AF4 mRNA transcript of the t(4;11)(q21;q23) was found in the girl with B-ALL and in patient with B-ALL after treatment. In patients with B-ALL we found MLL/AF10 cDNA segment specific for t(10;11)(p12;q23) by the mDx HemaVision kit.
Conclusions. On the basis of the concordance of the results it could be concluded that the method is suitable for the molecular genetic recognitions of leukemia. It offers the detection of many specific mRNA transcripts or cDNA segments of the chromosomal rearrangements simultaneously. We can obtain more information in shorter time and therefore reduce the costs of the test.
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References
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