Kelly Victoria R, Jones Susan P, Sammartino Holly L, Arocena Dennis Ian S, Madore Steven J
Molecular Biology Group, Coriell Institute for Medical Research , Camden, New Jersey.
Biopreserv Biobank. 2014 Jun;12(3):217-9. doi: 10.1089/bio.2013.0080.
Biorepository processing includes nucleic acid extractions in batch mode from a large number of blood samples from many different donors. Handling such a large number of biospecimens presents the challenge of ensuring that samples are not switched or mislabeled during processing. One approach for confirming donor identity from DNA samples is the use of multiplexed fluorescent PCR for detecting Short Tandem Repeat (STR) allelic-size polymorphisms for a set of common autosomal loci. While donor identity of DNA extracted directly from blood collected in standard tubes containing anticoagulants can be easily verified by generating STR profiles, RNA from blood collected in PAXgene Blood RNA tubes (PAXgene RNA tubes) is depleted of DNA and is not amenable to STR fingerprinting for donor identity verification. We investigated the feasibility of isolating DNA directly from blood collected in PAXgene RNA tubes for use as template for STR DNA fingerprinting for blood donor identity verification. We determined that DNA extraction can be performed manually with the QIAamp DNA Blood Minikit or on the QIAxtractor instrument with minimal pre-processing protocol additions, and that DNA isolated from blood collected in PAXgene RNA tubes is of sufficient quantity and quality for successful STR fingerprint analysis. Adaptation of quality assurance methods such as the PAXgene RNA tube DNA extraction/STR fingerprinting assay described here is a good practice that ensures that biobanking collections provide scientists with high quality, donor-verified biomaterial.
生物样本库处理包括以批量模式从许多不同捐赠者的大量血液样本中提取核酸。处理如此大量的生物样本带来了确保样本在处理过程中不被调换或错误标记的挑战。从DNA样本确认捐赠者身份的一种方法是使用多重荧光PCR检测一组常见常染色体位点的短串联重复（STR）等位基因大小多态性。虽然直接从含有抗凝剂的标准管中采集的血液中提取的DNA的捐赠者身份可以通过生成STR图谱轻松验证，但从PAXgene Blood RNA管（PAXgene RNA管）中采集的血液中的RNA不含DNA，不适合用于捐赠者身份验证的STR指纹识别。我们研究了直接从PAXgene RNA管中采集的血液中分离DNA作为STR DNA指纹识别模板用于献血者身份验证的可行性。我们确定，可以使用QIAamp DNA Blood Minikit手动进行DNA提取，或在QIAxtractor仪器上进行，只需添加最少的预处理方案，并且从PAXgene RNA管中采集的血液中分离的DNA在数量和质量上足以成功进行STR指纹分析。采用诸如本文所述的PAXgene RNA管DNA提取/STR指纹识别测定等质量保证方法是一种良好做法，可确保生物样本库为科学家提供高质量、经捐赠者验证的生物材料。
