DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT

<p>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 absen...

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Main Authors: Tadej Pajič, Elvira Ljutić, Peter Černelč
Format: Article
Language:English
Published: Slovenian Medical Association 2004-12-01
Series:Zdravniški Vestnik
Subjects:
Online Access:http://vestnik.szd.si/index.php/ZdravVest/article/view/2414
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spelling doaj-94c95edf429f4ee390d13f412aed839a2020-11-25T00:59:47ZengSlovenian Medical AssociationZdravniški Vestnik1318-03471581-02242004-12-017301883DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KITTadej Pajič0Elvira Ljutić1Peter Černelč2Enota za diagnostiko Klinični oddelek za hematologijo Klinični center Njegoševa 4 1525 Ljubljana SlovenijaEnota za diagnostiko Klinični oddelek za hematologijo Klinični center Njegoševa 4 1525 Ljubljana SlovenijaEnota za diagnostiko Klinični oddelek za hematologijo Klinični center Njegoševa 4 1525 Ljubljana Slovenija<p>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.</p><p>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.</p><p>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.</p><p>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.<br /><br /></p>http://vestnik.szd.si/index.php/ZdravVest/article/view/2414leukemiareverse transcriptionpolymerase chain reactionchromosomal rearrangementfusion genes
collection DOAJ
language English
format Article
sources DOAJ
author Tadej Pajič
Elvira Ljutić
Peter Černelč
spellingShingle Tadej Pajič
Elvira Ljutić
Peter Černelč
DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT
Zdravniški Vestnik
leukemia
reverse transcription
polymerase chain reaction
chromosomal rearrangement
fusion genes
author_facet Tadej Pajič
Elvira Ljutić
Peter Černelč
author_sort Tadej Pajič
title DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT
title_short DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT
title_full DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT
title_fullStr DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT
title_full_unstemmed DETECTION OF SPECIFIC CHROMOSOMAL REARRANGEMENT IN LEUKEMIA PATIENTS BY mDx HEMAVISION KIT
title_sort detection of specific chromosomal rearrangement in leukemia patients by mdx hemavision kit
publisher Slovenian Medical Association
series Zdravniški Vestnik
issn 1318-0347
1581-0224
publishDate 2004-12-01
description <p>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.</p><p>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.</p><p>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.</p><p>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.<br /><br /></p>
topic leukemia
reverse transcription
polymerase chain reaction
chromosomal rearrangement
fusion genes
url http://vestnik.szd.si/index.php/ZdravVest/article/view/2414
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