Mixed chimerism after sex-mismatched allogeneic BMT: evaluation of two molecular techniques.
Two different molecular techniques were used to monitor chimerism following 17 non-T cell-depleted BMTs from female donors to male recipients: pHY10, a Y chromosome-specific probe (Southern or slot blots), and a set of primers for Y chromosome sequence-specific amplification by the polymerase chain reaction (PCR). On Southern blots, male DNA was detectable at a level less than 1% of 10 micrograms DNA while cross-reactivity with autosomal sequences was avoided. On slot blots, male DNA was reliably detectable at levels less than 0.5%, even in small sample (0.5 microgram DNA). With the PCR technique, male DNA was detectable at levels of 1:10(6) to 1:10(7) of 0.5 microgram DNA. Slot blot and PCR results were concordant in 19 of 23 samples. Both techniques demonstrated a constant small mixed chimerism during the first year after BMT and in four of nine patients, this chimerism persisted even longer (up to 29 months after BMT).
Viard
F
F
Laboratoire d'Hématologie Université de Bordeaux II, France.
Merel
P
P
Bilhou-Nabera
C
C
Marit
G
G
Comeau
F
F
Gharbi
M J
MJ
Febrer
F
F
Belloc
F
F
Lacombe
F
F
Broustet
A
A
D000293:Adolescent; D000328:Adult; D015139:Blotting, Southern; D016026:Bone Marrow Transplantation / Q000473:pathology*; D002648:Child; D002675:Child, Preschool; D002678:Chimera / Q000235:genetics*; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D015336:Molecular Probe Techniques / Q000706:statistics & numerical data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D012723:Sex; D014019:Tissue Donors; D014998:Y Chromosome