The Assessment Of Microsatellite DNA Polym Orphism Similarity Between Donor And Recipient In HLA-matched Hematopoietic Stem Cell Transplantation

Objective:To evaluate the potential risk that the hematopoietic stem cell donor may suffer from the same disease as his/her HLA-matched recipient,by their similarity assessment of 19 specific short tandem repeat loci(CSF1PO,D21S11,D2S1338,D3S1358,D5S818,D6S1043,D7S820,D8S1179,D12S391,D13S317,D16S539,D18S51,D19S433,FGA,Penta D,Penta E,TH01,TPOX,vWA).Methods:Blood samples from 72 pairs of donors and recipients in HLA-matched hematopoietic stem cell transplantation were selected as matched group.Blood samples from 72 people,who have mismatched HLA(HLA-A,HLA-B and HLA-DR)compared to people in matched groups,were selected and made pairs with blood samples in matched groups,respectively,as mismatched group.HLA test results were provided by Dalian Blood Center.After the DNA extraction,19 specific short tandem repeat loci of each sample were tested by capillary electrophoresis.Four different fluorescently-labeled primers were employed for the specific amplification.FAM was used to label five loci(D19S433,D5S818,D21S11,D18S51 and D6S1043).HEX was used to label six loci(D3S1358,D13S317,D7S820,D16S539,CSF1PO and Penta D).TMR was used to label four loci(vWA,D8S1179,TPOX and Penta E).CAL Fluor Red 610 was used to label four loci(TH01,D12S391,D2S1338 and FGA4).After the amplification,electrophoresis analysis was performed by genetic analyzer and DNA Sequencer.Data were analyzed by software SPSS 11.5.X2 test was used to analyze the STR gene frequency,and calculate whether the gene loci matching of matched group had statistical significant difference(P<0.05)compared with that of mismatched group.Paired-samples T test was used to calculate whether total STR of matched group had statistical significant difference(P<0.05)compared with that of mismatched group.Results:The results of STR gene frequency in matched group,analyzed by X2 test,showed that there were significant differences(P<0.05)at gene loci D5S818(P=0.048),and TH01(P=0.015).The results of STRgene frequency between matched and mismatched groups,analyzed by X2 test,showed that there were significant differences(P<0.05)at gene loci D13S317(P=0.041),and D16S539(P=0.028).In the paired test,matching ratios of gene loci(D19S433、D2S1338、vWA、FGA、D5S818、TH01、CSF1PO、D18S51、D12S391、D16S539、D6S1043、D21S11、D13S317、D7S820)in matched group were higher than that in mismatched group,respectively.Extreme matching number of 19 STR gene loci in matched group was significant higher than that in mismatched group(P=0.006),respectively.However,when considering 19 gene loci as a system,and using individual as the unit to perform the comparison,there was no significant difference between two groups(P=0.251).Conclusion:The potential risk that the hematopoietic stem cell donor may suffer from the same disease as his/her HLA-matched recipient does exist in the statistical level,but the actual risk remains to be evaluated.