Rapid, Simple and Cost-Effective Detection of Human Cell Line Contamination

细胞系污染和错误识别在生物医学研究领域产生了巨大的影响， 这在很大程度上加剧了人们对不可复制结果的日益担忧，并对 全球生物医学研究预算。虽然在过去的几年里，人们对这些问题的普遍认识有所提高 多年来，诸如短串联重复(STR)简档的当前认证技术通常是劳动密集型的， 速度慢，成本高，不适合癌细胞系，这使得很难可靠和连续地监测 人类细胞培养。分析单核苷酸多态(SNPs)将为STR提供一个很好的替代方案 在监测癌细胞培养时，与STR标记相比，SNPs受基因组不稳定性的影响较小。 NuProbe的阻滞剂置换扩增(BDA)技术独特地实现了多重稀有等位基因 用聚合酶链式反应进行扩增。在NuProbe的BDA技术的基础上，我们建议开发一系列高度 检测种内交叉污染物的多重和高灵敏的基于qPCR的SNP图谱分析方法 在人类细胞培养中存在低水平(降至1%)。我们预计这些检测将转化为快速、 符合成本效益、可靠且易于使用的细胞系污染检测试剂盒，与常用的 定量聚合酶链式反应仪器。

Cell line contamination and misidentification have dramatic repercussions in the field biomedical research, largely contributing to the growing concerns about irreproducible results and posing a significant burden on the global biomedical research budget. While the general awareness about these issues has raised over the last years, current authentication technologies such as short tandem repeat (STR) profiling are often labor intensive, slow, costly and not suited for cancer cell lines, which make it very difficult to reliably and continuously monitor human cell cultures. Profiling single nucleotide polymorphisms (SNPs) would provide a good alternative to STR for monitoring cancer cell cultures, SNPs being less affected by genomic instabilities than STR markers. NuProbe’s blocker displacement amplification (BDA) technology uniquely enables multiplexed rare allele enrichment by PCR. Building upon NuProbe’s BDA technology, we propose to develop a series of highly multiplexed and highly sensitive qPCR-based SNP profiling assays to detect intra-species cross-contaminants present in human cell cultures at low levels (down to 1%). We anticipate these assays to translate into rapid, cost effective, reliable, and easy-to-use cell line contamination detection kits compatible with commonly available qPCR instruments.