PMID- 15527322
OWN - NLM
STAT- MEDLINE
DCOM- 20041223
LR  - 20191210
IS  - 1084-8592 (Print)
IS  - 1084-8592 (Linking)
VI  - 8
IP  - 2
DP  - 2004
TI  - Feasibility of a cost-effective approach to evaluate short tandem repeat markers 
      suitable for chimerism follow-up.
PG  - 87-91
AB  - BACKGROUND: Precise chimerism monitoring is important for the prediction of the 
      success of allogeneic bone marrow transplantation (BMT). Most of the current 
      procedures employed for chimerism follow-up with short tandem repeat (STR) 
      markers are either time-consuming, labor-intensive, or use expensive assays, 
      making it burdensome to perform large-scale studies of transplanted patients. 
      AIM: To set-up a simple nonradioactive method to investigate a set of STR markers 
      that could be used in the evaluation of chimerism status after allogeneic BMT. 
      METHOD: Six dinucleotide STRs (D2S123, D5S107, CRTL1, D7S500, D11S1356, and TP53) 
      were analyzed by touchdown (TD)-PCR followed by medium size non-denaturing 
      polyacrylamide gel electrophoresis and silver staining. The sensitivity of the 
      approach was evaluated by dilution competition assays. Peripheral blood samples 
      were taken from a group of 50 healthy Argentinean donors, two transplanted 
      patients, and their respective bone marrow donors. Buccal mucosa samples were 
      also obtained from the BMT recipients. RESULTS: Four markers, D2S123, D7S500, 
      D11S1356, and TP53, presented the highest heterozygosities (0.67-0.88) under our 
      experimental system. A sensitivity of 0.8-1.6% for chimerism detection was 
      consistently found for the different STR. The usefulness of these STR in 
      chimerism analysis was illustrated with the screening of related siblings 
      analyzing two transplanted patients with persistent mixed chimerism, which were 
      previously studied by fluorescence in situ hybridization (FISH). Similar 
      proportions of mixed chimerism were obtained with STR analysis compared with 
      those estimated by FISH. DISCUSSION: To our knowledge, this was the first study 
      of mixed chimerism using TD-PCR to achieve a highly specific STR amplification. 
      This approach allows simple and accurate chimerism quantification because it 
      avoids slippage of Taq polymerase on repeat stretches and prevents the 
      differential amplification of the shorter allele. STR heterozygosities and the 
      high level of sensitivity of this method demonstrated that this approach is not 
      only very informative in this population, but is also rapid (taking less than 14 
      hours) and cost-efficient. CONCLUSION: The data confirms that this method is a 
      useful tool applicable to routine large-scale STR genotyping and mixed chimerism 
      analysis in low-complexity laboratories worldwide.
FAU - Fundia, Ariela F
AU  - Fundia AF
AD  - Department of Genetics, Institute of Hematological Research, Mariano R Castex, 
      National Medicine Academy of Buenos Aires, Argentina. 
      affundia@hematologia.anm.edu.ar
FAU - De Brasi, Carlos
AU  - De Brasi C
FAU - Larripa, Irene
AU  - Larripa I
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Mol Diagn
JT  - Molecular diagnosis : a journal devoted to the understanding of human disease 
      through the clinical application of molecular biology
JID - 9614965
RN  - 0 (Genetic Markers)
SB  - IM
MH  - Bone Marrow Transplantation/physiology
MH  - Child
MH  - *Chimerism
MH  - Cost-Benefit Analysis
MH  - Female
MH  - Genetic Markers
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - Polymerase Chain Reaction/*economics/*methods
MH  - Sensitivity and Specificity
MH  - Tandem Repeat Sequences/*genetics
MH  - Transplantation Chimera/*genetics
EDAT- 2004/11/06 09:00
MHDA- 2004/12/24 09:00
CRDT- 2004/11/06 09:00
PHST- 2004/11/06 09:00 [pubmed]
PHST- 2004/12/24 09:00 [medline]
PHST- 2004/11/06 09:00 [entrez]
AID - 822 [pii]
AID - 10.1007/BF03260050 [doi]
PST - ppublish
SO  - Mol Diagn. 2004;8(2):87-91. doi: 10.1007/BF03260050.