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Homogeneous and digital proximity ligation assays for the detection of Clostridium difficile toxins A and B

Overview of attention for article published in Biomolecular Detection and Quantification, December 2016
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  • In the top 25% of all research outputs scored by Altmetric
  • High Attention Score compared to outputs of the same age (80th percentile)
  • Average Attention Score compared to outputs of the same age and source

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Title
Homogeneous and digital proximity ligation assays for the detection of Clostridium difficile toxins A and B
Published in
Biomolecular Detection and Quantification, December 2016
DOI 10.1016/j.bdq.2016.06.003
Pubmed ID
Authors

Harvinder S. Dhillon, Gemma Johnson, Mark Shannon, Christina Greenwood, Doug Roberts, Stephen Bustin

Abstract

The proximity ligation assay (PLA) detects proteins via their interaction with pairs of proximity probes, which are antibodies coupled to noncomplementary DNA oligonucleotides. The binding of both proximity probes to their epitopes on the target protein brings the oligonucleotides together, allowing them to be bridged by a third oligonucleotide with complementarity to the other two. This enables their ligation and the detection of the resulting amplicon by real-time quantitative PCR (qPCR), which acts as a surrogate marker for the protein of interest. Hence PLA has potential as a clinically relevant diagnostic tool for the detection of pathogens where nucleic acid based tests are inconclusive proof of infection. We prepared monoclonal and polyclonal proximity probes targeting Clostridium difficile toxins A (TcdA) and B (TcdB) and used hydrolysis probe-based qPCR and digital PCR (dPCR) assays to detect antibody/antigen interactions. The performance of the PLA assays was antibody-dependent but both TcdA and TcdB assays were more sensitive than comparable ELISAs in either single- or dualplex formats. Both PLAs could be performed using single monoclonal antibodies coupled to different oligonucleotides. Finally, we used dPCR to demonstrate its potential for accurate and reliable quantification of TcdA. PLA with either qPCR or dPCR readout have potential as new diagnostic applications for the detection of pathogens where nucleic acid based tests do not indicate viability or expression of toxins. Importantly, since it is not always necessary to use two different antibodies, the pool of potential antibodies useful for PLA diagnostic assays is usefully enhanced.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 44 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 44 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 11 25%
Student > Ph. D. Student 6 14%
Student > Master 6 14%
Student > Bachelor 4 9%
Other 4 9%
Other 6 14%
Unknown 7 16%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 10 23%
Agricultural and Biological Sciences 8 18%
Medicine and Dentistry 4 9%
Immunology and Microbiology 3 7%
Chemistry 3 7%
Other 8 18%
Unknown 8 18%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 8. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 12 July 2023.
All research outputs
#4,222,986
of 23,515,383 outputs
Outputs from Biomolecular Detection and Quantification
#30
of 71 outputs
Outputs of similar age
#80,503
of 419,752 outputs
Outputs of similar age from Biomolecular Detection and Quantification
#3
of 5 outputs
Altmetric has tracked 23,515,383 research outputs across all sources so far. Compared to these this one has done well and is in the 81st percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 71 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 12.0. This one has gotten more attention than average, scoring higher than 57% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 419,752 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 80% of its contemporaries.
We're also able to compare this research output to 5 others from the same source and published within six weeks on either side of this one. This one has scored higher than 2 of them.