Author/Editor     Pajič, Tadej; Ljutić, Elvira; Černelč, Peter
Title     Prepoznavanje značilnih kromosomskih prireditev z reagenčnim kompletom mDx HemaVision pri bolnikih z levkemijo
Translated title     Detection of specific chromosomal rearrangement in leukemia patients by mDx HemaVision kit
Type     članek
Source     Zdrav Vestn
Vol. and No.     Letnik 73, št. Suppl 1
Publication year     2004
Volume     str. I-9-12
Language     slo
Abstract     Background. The mDx HemaVision kit is a qualitative multiplex and nested Reverse Transcription. Polymerase Chain Reaction (RTPCR) test designed to detect 28 different translocations or chromosomal rearrangement, found to be specific for particular subtypes of leukemia. The presen.ce 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 sarnples 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)(q22;q22) 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 bifenotypic 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 CBFbeta/MYH11 mRNA transcript specific for invl6(p13;q22) was obtained in AML patient with abnormal eozinophiles in bone marrow. MLL/AF4 mRNA transcript of the t(4;11)(q21;q23) was found in the girl with BALL and in patient with B-ALL after treatment. (Abstract truncated at 2000 characters).
Summary     Izhodišča. Reagenčni komplet mDx HemaVision je namenjen za kvalitativno ugotavljanje 28 različnih kromosomskih preureditev, značilnih za posamezne podvrste levkemij z načinom obratnega prepisovanja, hkratne in vgnezdene verižne reakcije s polimerazo. S to reakcijo lahko določimo prisotnost ali odsotnost specifičnih mRNK prepisov oziroma cDNK segmentov po obratnem prepisovanju združenih ali okvarjenih genov, ki nastanejo zaradi kromosomskih preureditev. Bolniki in metode. Uporabnost reagenčnega kompleta smo preverili v 26 vzorcih RNK bolnikov z akutno levkemijo in 7 s kronično mieloproliferativno boleznijo ter primerjali izsledke med reagenčnim kompletom mDx HemaVision in usklajenim protokolom RT-PCR pri ugotavljanju značilnih prepisov mRNK t(9;22)(q34;q11), t(8;21)(q22;q22), inv16(p13;q22), t(15;17)(q21;q21) in t(4;11)(q21;q23). RNK smo pridobili iz enojedrnih celic kostnega mozga po gostotnem gradientnem centrifugiranju preko fikola in z uporabo reagenčnega kompleta High Pure RNA Isolation. Sintezo komplementarne DNK in verižno reakcijo s polimerazo smo izvedli, kot je opisano v protokolu reagenčnega kompleta ali mDx HemaVision v usklajenem protokolu RT-PCR. Pridelke PCR reakcij smo ocenjevali z agarozno gelsko elektroforezo, barvanjem gelov z etidijevim bromidom in detekcijo pod UV lučjo. Rezultati. Dobili smo 100-odstotno ujemanje izsledkov za preiskave, ki so skupne obema postopkoma. Značilen BCRABL mRNK prepis t(9;22)(q34;q11) smo dokazali pri bolniku z BALL, bolniku z bifenotipsko levkemijo in pri štirih bolnikih s KML. AHL1-ETO mRNK prepis t(8;22)(q22;q22) smo ugotovili pri dveh bolnikih z AML. Pri enem bolniku z AML z nenormalnimi eozinofilci v kostnem mozgu smo dokazali CBFbeta/ MYH11 mRNK prepis inv16(p13;q22). MLL/AF4 mRNK prepis t(4,11))(q21;q23) smo ugotovili pri deklici z B-ALL in bolniku z BALL po zdravljenju s citostatiki. (Izvleček skrajšan pri 2000 znakih).
Descriptors     LEUKEMIA
CHROMOSOME ABERRATIONS
REAGENT KITS, DIAGNOSTIC
POLYMERASE CHAIN REACTION
ELECTROPHORESIS, AGAR GEL
LEUKEMIA, LYMPHOCYTIC, ACUTE
LEUKEMIA, B-CELL, ACUTE
LEUKEMIA, T-CELL, ACUTE
LEUKEMIA, MYELOID, CHRONIC