应用微卫星标志进行脐血干细胞移植后的植活检测

Monitoring of engraftment with microsatellite markers following cord blood stem cell transplantation

目的 :应用微卫星标志进行脐血干细胞移植后的植活检测 ,动态检测植活类型。方法 :对 12例脐血移植患者进行植活检测 ,其中 10例作动态检测 ,另 2例作随访检测。选用D12s2 2 5 7、D12s2 398、Dxs991、Dxs1199、D17s12 92、D5s8186个微卫星位点为引物进行PCR扩增 ,扩增产物经 8%聚丙烯酰胺凝胶电泳作银染色分析。取供者脐血或外周血及受者口腔黏膜DNA做为对照。结果 :10例动态检测者移植后 14d ,6例为混合性嵌合(MC) ;第 2 1天 ,7例为MC或完全性嵌合 (CC) ,其中 4例为CC ;第 2 8天 ,6例为CC。 2例植活失败的病例始终未检测出嵌合表达。 2例随访检测结果为CC。结论 :应用微卫星标志进行脐血移植后的植活检测 ,能早期检测是否植活及嵌合类型 。

Objective:To monitor engrafment and consecutive hematopoietic chimerism(HC) with microsatellite markers following cord blood transplantation(CBSCT).Method:Among 12 cases , Samples of BM were collected at certain times in 10 cases after CBSCT ,other 2 cases were followed up to detect HC .DNA removed from donors peripheral blood and cord blood , mouth mocous memberance of recipients to be tested as control. 6 microsatellite site D12s2257,D12s2398,Dxs991,Dxs1199,D17s1292,D5s818 were selected as primer to perform PCR Amplifiction. Amplification products were analysed by electrophoresis on 8% ployacrylamid gel and silver stained.Result:In 10 cases, the consecutive result were detcted: mixer chimerism(MC) occured in 6 cases at +14 day;MC in 3cases and complete chimerism(CC) in 4 cases at +21 day; 6 cases were all CC at +28 day and +90 day. 2 cases were failed in CBSCT which did not detected HC at all times. As for other 2 cases , the type of HC was CC.Conclusion:Monitoring of engrament with microsatellite markers following cord blood transplantation can be applied in detection of implant in early stage. The method can provide warrant evidence of successful engraft ment for clinical treatment.