摘要
应用四条引物（Ｒ＿５，Ｒ＿６，Ｄ＿３，Ｄ＿４）通过逆转录聚合酶链反应（ＲＴ－ＰＣＲ）检测了２０例初治急性早幼粒细胞白血病（ＡＰＬ）患者的ＲＡＲα／ＭＹＬ融合ｍＲＮＡ。建立的方法可在０．１ｎｇ细胞总ＲＮＡ中检出ＲＡＲα／ＭＹＬ融合ｍＲＮＡ，相当于在１０￣４～１０￣５个正常细胞中检出一个ＡＰＬ细胞，扩增产物的特异性经ＫｐｎＩ酶切法确认。１８例患者表达Ｂ型融合ｍＲＮＡ（２９０ｂｎ片段），２例表达Ａ型（３１１ｂｐ片段）。２例患者在完全缓解（ＣＲ）达４周和１６周时仍可检出融合ｍＲＮＡ。１例患者在ＣＲ期ＲＴ－ＰＣＲ检测持续阳性，４周后；临床复发。４例在ＣＲ期随访９～２９周融合ｍＲＮＡ均阴性。２例急性白血病在维甲酸治疗前拟诊ＡＰＬ，但ＲＴ。ＰＣＲ检测阴性，这２例对维甲酸治疗无效，核型分别是ｔ（８；２１）和正常。我们认为，ＲＡＲα／ＭＹＬ融合ｍＲＮＡ与ＭＹＬ／ＲＡＲα融合ｍＲＮＡ一样，也是诊断和监测ＡＰＬ的一个敏感和特异的标志。
he RARα/MYL fusion transcripts were determinedin bone marrow samples from2 0 cases of de novo acutepromyelocytic leukemia(APL)by reverse transcriptase-polymerase chain reaction(RT-PCR)technique. Fourprimers(R5 ,R6;D3;D4)were u;ed in this study.Thetest was able to detect the amplified R ARα/MYL fusiontranscripts in the presence of 0.1lng total RNA,whichrepresented cell dilution in 10￣(-4)～10￣(-5).The specificityof amplified products could be further identified by Kpn Icleavage test. Ninety percent(1 8/20)cases expressedtype B form (290 bp fragment) and the other two casesbelonged to type A form(311 bp fragment).Seven ofthe 20 cases were able to be followed up after treatrnent.The fusion mRNA was still detected in two patients whohad been in cornplete remission(CR) for 4 and 16 weeksrespectively. One patient showed continuouslv RT-PCRpo;itive during CR and relap:ed morphologically fourweeks later. Other 4 patietns in CR were RT-PCR neg-ative during 9～29 weeks follow-up.Two caseswith acute myeloid leukemid, were classifiedasFAB-M_3before retinoid acid treatment,but their RT-PCR results were negative. Both of thern failed in thetreatment,and their karyotype showed typical t ( 8; 21)and normal respectively. We concluded that detection ofthe RARα/MYL fusion mRNA was compatible with MYL/RARα fusion mRNA,it was also a senshive andspecific marker for diagnosis and monitoring of minimalresidual disease in APL.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
1994年第2期57-63,共7页
Chinese Journal of Hematology
基金
卫生部科研基金
关键词
白血病
聚合酶链反应
诊断
RARα/MYL fusion mRNA Acute promyelocytic leukemia Polymerase chain reaction Minimal residual dis-ease
