Johnson Donald J, Martin Liane R, Roberts Katherine A
School of Criminal Justice and Criminalistics, California State University, Los Angeles, College of Health and Human Services, 90032-8163, USA.
J Forensic Sci. 2005 Jul;50(4):802-8.
The purpose of this study was to compare the effectiveness of the QIAGEN QIAamp Stool Mini Kit against a standard phenolchloroform procedure for the extraction, quantitation, and STR-typing of human nuclear DNA from human feces. Stools from six subjects were sampled by swabbing and excision. Samples extracted with the QIAamp kit gave a wide range of DNA yields, whereas those extracted by the organic method yielded no DNA. DNA was not recovered from one subject's stools by either procedure. The QIAamp extracts were amplified with the Profiler Plus and COfiler kits, and PCR inhibition was observed with DNA extracts that were further concentrated. Substitution of water or TE-4 for the QIAamp elution buffer eliminated most, if not all, of the inhibition. A modified QIAamp procedure was used to extract thirty samples, which were subjected to one of five environmental conditions. DNA was recovered from all of these samples, and typing results were obtained on 93% of the samples.
本研究的目的是比较QIAGEN QIAamp粪便迷你试剂盒与标准酚氯仿法从人粪便中提取、定量和进行人核DNA短串联重复序列(STR)分型的有效性。通过擦拭和切除的方式采集了6名受试者的粪便样本。用QIAamp试剂盒提取的样本得到了广泛的DNA产量范围,而通过有机方法提取的样本未获得DNA。两种方法均未从一名受试者的粪便中回收DNA。用Profiler Plus和COfiler试剂盒对QIAamp提取物进行扩增,发现进一步浓缩的DNA提取物存在PCR抑制现象。用水或TE-4替代QIAamp洗脱缓冲液消除了大部分(如果不是全部)抑制作用。采用改良的QIAamp方法提取了30个样本,这些样本处于五种环境条件之一。所有这些样本均回收了DNA,93%的样本获得了分型结果。