Standardisation of multiplex fluorescent short tandem repeat analysis for chimerism testing
Standardisation of multiplex fluorescent short tandem repeat analysis for chimerism testing
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DOI：
10.1038/sj.bmt.1703162
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发表时间：
2001-09-01
影响因子：
4.8
通讯作者：
Criel, A
中科院分区：
文献类型：
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作者：
Nollet, F;Billiet, J;Criel, A
为了评估异基因造血干细胞移植后细胞的来源，我们优化并评估了两种基于多重荧光短串联重复序列（STR）分析的市售系统（AmpFlSTR Profiler Plus和GenePrint Powerplex-16）。一个标准的程序解释电泳被认为是必不可少的，以获得重现性的结果。基于供体和受体等位基因的相对长度，区分I型（没有共享等位基因用于计算嵌合性）、II型（一个共享等位基因和一个非共享等位基因用于计算嵌合性）或III型（无信息）等位基因分布类型。此外，断续峰被认为是排除用于分析的标志物的重要标准。AmpFlSTR Profiler Plus系统的实验室内和多中心评估表明，可以测定混合血液样本，绝对偏差小于2%。对于I型标记物，灵敏度阈值设定为5%，对于II型标记物，灵敏度阈值设定为10%，因为在低嵌合性值时相对不精密度增加。两个系统共有的STR标记的嵌合率计算值之间没有显着差异。通过对26例异基因外周血干细胞移植的监测，证明了该方法的适用性。
To evaluate the origin of cells after allogeneic haematopoietic stem cell transplantation we optimised and evaluated two commercially available systems (AmpFlSTR Profiler Plus and GenePrint Powerplex-16) which are based on multiplex fluorescent short tandem repeat (STR) analysis. A standard procedure for interpretation of electropherographs was found essential to obtain reproducible results. On the basis of the relative length of donor and recipient alleles, TYPE-I (no shared alleles are used to calculate chimerism), TYPE-II (one shared and one unshared allele is used to calculate chimerism) or TYPE-III (not informative) allelic distribution types were distinguished. Also, stutter peaks were recognised as an important criterion to exclude a marker for analysis. Intralaboratory and multicentre evaluation of the AmpFlSTR Profiler Plus system showed that mixed blood samples could be determined with an absolute deviation of less than 2%. A sensitivity threshold was set at 5% for TYPE-I and 10% for TYPE-II markers since relative imprecision increases at low chimerism values. No significant difference of calculated chimerism values was observed between STR markers shared between both systems. By monitoring 26 allogeneic peripheral blood stem cell transplants, the applicability of the proposed method was demonstrated.