Improving precision of proximity ligation assay by amplified single molecule detection

Rongqin Ke; Rachel Yuan Nong; Simon Fredriksson; Ulf Landegren; Mats Nilsson

doi:10.1371/journal.pone.0069813

Improving precision of proximity ligation assay by amplified single molecule detection

PLoS One. 2013 Jul 16;8(7):e69813. doi: 10.1371/journal.pone.0069813. Print 2013.

Authors

Rongqin Ke¹, Rachel Yuan Nong, Simon Fredriksson, Ulf Landegren, Mats Nilsson

Affiliation

¹ Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Solna, Sweden.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biological Assay / methods*
Real-Time Polymerase Chain Reaction / methods

Grants and funding

The work was funded by grants from the Knut and Alice Wallenberg foundation, the Swedish Research Council, and the European Community’s 7th Framework Program (FP7/2007-2013) under grant agreement n°259796 (DiaTools). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.