利用FISH及PCR法研究肝和肾移植受者的血液嵌合性

Using FISH and PCR techniques to detect microchimerism in liver and renal transplant recipients

目的 证实肝、肾移植受者末稍血中存在供者细胞 ,证明血液嵌合性的成立。方法 女性活体肝移植受者 9例、肾移植受者 3 8例。供者均为男性。利用FISH法检测Y染色体 ;利用PCR及nestedPCR法检测Y染色体上特异的基因片断SRY和DYZ 1。结果 使用FISH法 ,肝、肾移植受者均未发现Y染色体的存在。利用PCR法检测 ,9例肝移植受者Y染色体特异的基因片断SRY和DYZ 1均为阳性。肾移植受者SRY阳性 3 2例 (84.2 % ) ,阴性 6例 (15 .8% ) ;DYZ 1阳性 3 5例(92 .1% ) ,阴性 3例 (7.9% )。结论 长期存活的肝、肾移植受者 ,血液嵌合性是成立的 ,且出现频率差异无显著性。FISH法检测末稍血中的Y染色体是困难的 。

Objective To detect cells of donor origin in the peripheral blood of recipients of liver transplant and renal transplants to prove the presence of microchimerism. Methods Y chromosome was detected using fluorescence in situ hybridization (FISH) technique. SRY and DYZ-1 region of the Y chromosome was detected using both PCR and nested PCR techniques. Results Y chromosome was not detected in both liver transplant and renal transplant recipients by FISH technique. Nine cases of Liver transplant recipients were detected positive for SRY and DYZ-1 region of the Y chromosome by PCR technique. SRY region was detected positive in 32 ( 84.2?%) of 38 renal transplant recipients, negative in 6 ( 15.8?%) of 38; DYZ-1 region positive in 35 ( 92.1?%) of 38, negative in 3 ( 7.9?%) of 38. Conclusions Microchimerism existed in both liver and renal transplant recipients with long term survival after transplantation, and there was no significant difference in detectable frequency of microchimerism between liver and renal transplant recipients. Detecting Y chromosome in peripheral blood is difficult with FISH assay.