Improving precision of proximity ligation assay by amplified single molecule detection.

Improving precision of proximity ligation assay by amplified single molecule detection.
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DOI:
10.1371/journal.pone.0069813
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Nilsson M
Nilsson M
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Ke R;Nong RY;Fredriksson S;Landegren U;Nilsson M

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邻位连接试验(PLA)已被证明是一种稳健的蛋白质检测方法。该技术的特点是高灵敏度和特异性,但检测精度可能受到PCR读数的限制。为了研究这种潜在的局限性并提高精确度,我们开发了一种基于扩增单分子检测的用于液体中蛋白质测量的数字邻近连接测定法。该测定法显示出显着改善的精度,从而也检测灵敏度,在传统的实时PCR读出。
Proximity ligation assay (PLA) has been proven to be a robust protein detection method. The technique is characterized by high sensitivity and specificity, but the assay precision is probably limited by the PCR readout. To investigate this potential limitation and to improve precision, we developed a digital proximity ligation assay for protein measurement in fluids based on amplified single molecule detection. The assay showed significant improvements in precision, and thereby also detection sensitivity, over the conventional real-time PCR readout.
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