The Establishment And Contrast Research Of Two Different Methods For Chimerism Testing After Allogeneic Stem Cell Transplantation

Allogeneic stem cell transplantation is the main treatment of choice for many hematological malignancies, as well as some nonmalignant hematological disorders, but disease relapse is still the most significant obstacle to allo-HSCT. Successful transplantation is a complex process, it is important to distinguish between host and donor origin of hematopoietic cells in order to monitor the engraftment process, relapse and intervention immunotherapy. There are many technologies to detect chimerism, the most useful method is PCR method based on the DNA polymorphisms. Now, the methods with short tandem repeats and single nucleotide polymorphisms have becoming the hot field. The method of STR with fluorescence has become the main testing method in abroad, but our country still use the method of STR silver-staining. The contrast researches about the sensitivity and accuracy of these two methods are seldom. Our research just based on it, we established the two different methods and compared the sensitivity and accuracy to evaluate the reliability of testing results. The goal was to service better for clinic.Methods: (1) To establish the method of STR silver-staining for testing chimerism .We choose 6 STRs with high polymorphisms from the CODIS data bank of American, then designed and synthesized the primer. The PCR amplificationproduct of 6 STRs was stained with silver after denaturation polyacrylamide gel electrophoresis. Signals were analyzed by imaging system. (2) To investigate chimeric status of 46 patients after allogeneic stem cell transplantation in General Hospital of China PLA between September 2002 and January 2005. (3) To establish the method of STR with fluorescence and capillary electrophoresis. AmpFSTR Profiler Plus? ID PCR Amplification Kit of ABI company were utilized and amplification products were run capillary electrophoresis by ABI...