Quantification of chimerism within peripheral blood, bone marrow and purified leukocyte subsets: comparison of singleplex and multiplex PCR amplification of short tandem repeat (STR) loci
Quantification of chimerism within peripheral blood, bone marrow and purified leukocyte subsets: comparison of singleplex and multiplex PCR amplification of short tandem repeat (STR) loci
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DOI：
10.1111/j.1600-0609.2005.00588.x
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发表时间：
2006-03-01
影响因子：
3.1
通讯作者：
Koehl, U
中科院分区：
文献类型：
--
作者：
Beck, O;Seidl, C;Koehl, U
目的和方法：嵌合分析已成为造血同种异体干细胞移植后的常规诊断程序，用于早期检测疾病复发或移植失败。虽然一些中心开发了单独的内部短串联重复 (STR) 系统，但其他中心更喜欢商业多重 PCR 系统。然而，人们对测定间变异知之甚少，这可能对治疗决策产生重大影响。因此，我们使用不同的样本来源（例如外周血 (PB)、骨髓 (BM) 和特定白细胞亚群）将两种商业多重 PCR 试剂盒与我们的内部 STR 系统进行了比较。 结果：分析了 18 名儿科患者的 50 个样本。对于PB、BM 和白细胞亚型这两种材料，所测试的STR 系统之间均未观察到显着差异。作为内部和商业 STR 系统组成部分的每个 STR 引物的嵌合分析没有显示显着差异。结论：我们的分析表明，即使使用不同的样品来源，两种检测也可以获得相似的结果。对不同测试系统的进一步评估将有助于提高早期临床干预嵌合分析的实验室间标准化。
Objective and Methods: Chimerism analysis has become a routine diagnostic procedure after haematopoietic allogeneic stem cell transplantation for early detection of relapse of disease or graft failure. Whereas some centres developed individual in-house short tandem repeat (STR) systems, others prefer commercial multiplex PCR systems. However, little is known about inter-assay variation, which could have a significant impact on treatment decision. We therefore compared two commercial multiplex PCR kits with our in-house STR system using different sample sources, such as peripheral blood (PB), bone marrow (BM) and specific leukocyte subsets.Results: Fifty samples of eighteen paediatric patients were analysed. For neither material, PB, BM and leukocyte subtypes, a significant difference between the STR systems tested was observed. Chimerism analyses of each single STR primer, which is component of both the in-house and the commercial STR system, did not reveal significant differences.Conclusion: Our analysis demonstrates that similar results can be obtained with both assays, even when using various sample sources. Further evaluation of different test systems will help to increase interlaboratory standardisation of chimerism analyses for early clinical intervention.