Martinelli G, Trabetti E, Zaccaria A, Farabegoli P, Buzzi M, Testoni N, Calori E, Bandini G, Rosti G, Belardinelli A
Institute of Haematology L. and A. Seragnoli, University of Bologna, Italy.
Bone Marrow Transplant. 1993 Aug;12(2):115-20.
The role of mixed hematopoietic chimerism in engraftment and relapse after allogeneic BMT remains unclear. To better evaluate post-transplant chimerism we used polymerase chain reaction (PCR) in vitro amplification of four single locus simple repetitive DNA sequences, all of which vary extensively in their repeat number among different individuals: variable number of tandem repeats D1S80, APOB and D17S5, and the tetranucleotide repeat F8VWF. We tested 13 cases of CML, four of multiple myeloma (MM), three of ANLL and one of B-CLL. In a sequential analysis protocol with the different loci, the donor could be distinguished from the recipient in 14 of 20 (70%) pairs with the first marker used (D1S80). When a donor of opposite sex was involved, karyotyping and Y chromosome-specific PCR were also used. With the use of the four markers, chimerism was identified in all the pairs. Mixed chimerism was present in 5 patients, and complete chimerism in 15. No patients relapsed. The application of PCR for documenting post-transplant chimerism has several advantages over Southern blotting: increased sensitivity, use of small amounts of sample, ease of preparation of DNA, elimination of restriction enzyme analysis and of radioisotopes, and speed.
混合造血嵌合体在异基因骨髓移植后的植入及复发中的作用仍不清楚。为了更好地评估移植后的嵌合体状态，我们采用聚合酶链反应（PCR）对四个单基因座简单重复DNA序列进行体外扩增，所有这些序列在不同个体间的重复次数差异很大：串联重复序列D1S80、载脂蛋白B（APOB）和D17S5，以及四核苷酸重复序列F8VWF。我们检测了13例慢性粒细胞白血病（CML）、4例多发性骨髓瘤（MM）、3例急性非淋巴细胞白血病（ANLL）和1例B细胞慢性淋巴细胞白血病（B-CLL）。在对不同基因座进行序列分析的方案中，使用第一个标记物（D1S80）时，在20对样本中有14对（70%）能够区分供体和受体。当涉及异性供体时，还采用了核型分析和Y染色体特异性PCR。使用这四种标记物，所有样本对中均检测到嵌合体。5例患者存在混合嵌合体，15例为完全嵌合体。无患者复发。与Southern印迹法相比，应用PCR记录移植后嵌合体具有几个优点：灵敏度提高、使用少量样本、DNA制备简便、无需限制性酶切分析和放射性同位素，且速度快。
