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Improved amplification efficiency on stool samples by addition of spermidine and its use for non-invasive detection of colorectal cancer

机译：通过添加亚精胺提高粪便样品的扩增效率，并将其用于大肠癌的非侵入性检测

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BackgroundududUsing quantitative methylation-specific PCR (QM-MSP) is a promising method for colorectal cancer (CRC) diagnosis from stool samples. Difficulty in eliminating PCR inhibitors of this body fluid has been extensively reported. Here, spermidine is presented as PCR facilitator for the detection of stool DNA methylation biomarkers using QM-MSP. We examined its effectiveness with NPY, PENK and WIF1, three biomarkers which we have previously shown to be of relevance to CRC.udResultsududWe determined an optimal window for the amplification of the albumin (Alb) gene (100 ng of bisulfite-treated stool DNA added of 1 mM spermidine) at which we report that spermidine acts as a PCR facilitator (AE = 1680%) for SG RT-PCR. We show that the amplification of methylated PENK, NPY and WIF1 is considerably facilitated by QM-MSP as measured by an increase of CMI (Cumulative Methylation Index, i.e. the sum of the three methylation values) by a factor of 1.5 to 23 fold in individual samples, and of 10 fold in a pool of five samples.udConclusionsududWe contend that spermidine greatly reduces the problems of PCR inhibition in stool samples. This observed feature, after validation on a larger sampling, could be used in the development of stool-based CRC diagnosis tests.
机译：背景 ud ud使用定量甲基化特异性PCR（QM-MSP）是一种从粪便样本诊断大肠癌（CRC）的有前途的方法。已经广泛报道了消除该体液的PCR抑制剂的困难。在这里，亚精胺作为PCR促进剂，用于使用QM-MSP检测粪便DNA甲基化生物标志物。我们用NPY，PENK和WIF1这三种生物标志物检查了它的有效性，这些生物标志物先前已证明与CRC相关。 -处理的粪便DNA中加入了1 mM亚精胺），在该处我们报道亚精胺起了SG RT-PCR的PCR促进剂作用（AE = 1680％）。我们显示，QM-MSP极大地促进了甲基化PENK，NPY和WIF1的扩增，这是通过CMI（累积甲基化指数，即三个甲基化值的总和）增加1.5到23倍来衡量的 ud结论 ud ud我们认为亚精胺大大减少了粪便样品中PCR抑制的问题。经过大量抽样验证后，此观察到的特征可用于开发基于粪便的CRC诊断测试。

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