Nuckols J D, Rasheed B K, McGlennen R C, Bigner S H, Stenzel T T
Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA.
Am J Clin Pathol. 2000 Jan;113(1):135-40. doi: 10.1309/QP7P-J49V-8Q15-36MT.
Several methods have been used to evaluate engraftment after allogeneic bone marrow transplantation (BMT). We assessed the usefulness of a multiple short tandem repeat (STR) amplification kit combined with a capillary electrophoresis unit for DNA identity analysis in the evaluation of engraftment after BMT. For 17 of 18 patients, at least 1 locus showed unique alleles for the donor and the recipient. In all cases, at least 1 locus was informative for the presence of small amounts of recipient DNA. The results from STR analysis were the same as Southern blot analysis in 14 of 17 cases. Differences included mixed chimerism detected only with STR analysis, informative loci present only with STR analysis, and informative loci present only with Southern blot analysis (1 case each). By using mock mixed chimeras, minor populations of 5% were detected routinely in all loci using the kit manufacturer's default protocol. By increasing the amount of amplified DNA, minor populations of 1% were detected in all cases but not in all loci. This single reaction technique provides for faster results, reduced workforce needs, and greater sensitivity than traditional Southern blot.
已经使用了几种方法来评估异基因骨髓移植(BMT)后的植入情况。我们评估了一种多重短串联重复序列(STR)扩增试剂盒与毛细管电泳单元相结合用于DNA同一性分析在BMT后植入评估中的实用性。对于18例患者中的17例,至少有1个位点显示供体和受体的独特等位基因。在所有情况下,至少有1个位点对于少量受体DNA的存在具有信息价值。STR分析结果与17例中的14例Southern印迹分析结果相同。差异包括仅通过STR分析检测到的混合嵌合体、仅通过STR分析存在的信息位点以及仅通过Southern印迹分析存在的信息位点(各1例)。通过使用模拟混合嵌合体,使用试剂盒制造商的默认方案在所有位点常规检测到5%的少数群体。通过增加扩增DNA的量,在所有情况下均检测到1%的少数群体,但并非在所有位点。这种单一反应技术比传统的Southern印迹提供了更快的结果、减少了劳动力需求并且具有更高的灵敏度。