PCR－小卫星DNA指纹图在异基因造血干细胞移植植活指标监测中的应用

为了解异基因骨髓移植治疗某些遗传及恶性血液性疾病的植活监测指标，用聚合酶链式反应（ＰＣＲ）扩增具有高度多态性的小卫星ＤＮＡ３３．１、３３．６位点，联合地高辛标记的位点特异性寡核苷酸探针杂交的方法，以小卫星ＤＮＡ指纹图为特异性遗传标志，对６例异基因骨髓移植，１例脐血移植进行植活指标监测，６例获得供者源性细胞植活证据。结果显示：小卫星ＤＮＡ指纹图个体特异性强，可作为监测异基因造血干细胞移植植活的可靠格标，尤其是同血型，同性别，ＨＬＡ配型表型全相合的移植；该方法程序简单，操作方便，灵敏度高，可达０．５％，最早获得植入证据的时间是＋１５天，仅需模板ＤＮＡ１００ｎｇ；如果扩增位点增加，可提高个体识别能力。

ENGRAFIMENT AFTER ALLOGENEIC BONE MARROW TRANSPLANTATION MONITORED WITH DNA FINGERPRINTING OF HUMAN MINISATELLITE DNA AMPLIFIED BY POLYMERASE CHAIN REACTION

We developed a technique of amplifying human hypervariable minisatellite DNA by PCR combined with digoxigenin-labeled locus-specific oligonucleotide hybridization. DNA fingerprinting by this technique was used as a specific genetic marker to monitor the engraftment after BMT in six patients and after umbilical cord blood transplantation in one patient.Six of them gained evidence of engraftment.The results showed that the minisatellite DNA fingerprint is of high specificity and is valuable in confirming engraftment after BMT,especially when the patient and the donor are HLA identical,of the same sex,and have the same ABO-Rh blood type. The advantages of this technique are that there is no contamination by radioisotopes,and it is not restricted by the half-life. It is simple and bighly sensitive,Engraftment of donor marrow can be determined as early as 15 day posttransplantation using 100g of DNA. The ability of identifying individuals can be enhanced by increasing the number of amplified minisalellite DNA loci.