[Short tandem repeat panel settled for quantitative chimerism analysis following allogeneic stem cell transplantation and its application].
To evaluate the roles of 8 short tandem repeats (STR) loci as STR panel in quantitative analysis of chimerism following transplantation, the primers were synthesized and marked with different dyes for D3S3045, D4S2366, D4S2639, D5S818, D13S317, D18S1002, D20S481 and D22S689. The blood samples of 15 cases received allogeneic stem cell transplantation were collected before and after transplantation, then DNA was extracted and amplified with these primers, and was further analysed under ABI Genetic Analyser 3100 to select suitable informative STR locus. Donor/recipient dilution series were prepared to get standard curves in selected loci, the DNAs extracted at different days after transplantation were used to quantitatively analyze the chimerism in patients according to the values of peak area or peak height of fluorescent signals. The standard curves can be used to calculate the chimerism by plotting the respective R/D quotient value against the percentage of recipient DNA. The results indicated that the calculated chimerism was in concordance with the donor/recipient dilution. The STR panel succeeded in identifying at least one informative marker and quantitative monitoring the chimerism after HSCT in 15 donor-recipient pairs and a relapsed case was diagnosed. It is concluded that the STR panel and its detection method can accurately and quantitatively monitor the chimerism after allogeneic HSCT, which is more economical and flexible than using commercial kits.
Yang
Ying
Y
Shanghai Institute of Blood Transfusion, Shanghai Blood Center, Shanghai, China. yangyinghla@yahoo.com.cn
Wu
Wen
W
Wang
Ling-Ling
LL
Chen
He-Ping
HP
Zhu
Zi-Yan
ZY
D004247:DNA / Q000235:genetics; D017931:DNA Primers; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D018895:Microsatellite Repeats; D018183:Transplantation Chimera / Q000235:genetics*
D017931:DNA Primers; D004247:DNA