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Development of a rapid recombinase polymerase amplification assay for the detection of Streptococcus pneumoniae in whole blood

Overview of attention for article published in BMC Infectious Diseases, October 2015
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Title
Development of a rapid recombinase polymerase amplification assay for the detection of Streptococcus pneumoniae in whole blood
Published in
BMC Infectious Diseases, October 2015
DOI 10.1186/s12879-015-1212-5
Pubmed ID
Authors

Eoin Clancy, Owen Higgins, Matthew S. Forrest, Teck Wee Boo, Martin Cormican, Thomas Barry, Olaf Piepenburg, Terry J. Smith

Abstract

Streptococcus pneumoniae is an important cause of microbial disease in humans. The introduction of multivalent vaccines has coincided with a dramatic decrease in the number of pneumococcal-related deaths. In spite of this, at a global level, pneumococcal infection remains an important cause of death among children under 5 years of age and in adults 65 years of age or older. In order to properly manage patients and control the spread of infection, a rapid and highly sensitive diagnostic method is needed for routine implementation, especially in resource-limited regions where pneumococcal disease is most prevalent. Using the gene encoding leader peptidase A as a molecular diagnostics target, a real-time RPA assay was designed and optimised for the detection of S. pneumoniae in whole blood. The performance of the assay was compared to real-time PCR in terms of its analytical limit of detection and specificity. The inhibitory effect of human genomic DNA on amplification was investigated. The potential clinical utility of the assay was investigated using a small number of clinical samples. The RPA assay has a limit of detection equivalent to PCR (4.0 and 5.1 genome equivalents per reaction, respectively) and was capable of detecting the equivalent of <1 colony forming unit of S. pneumoniae when spiked into human whole blood. The RPA assay was 100 % inclusive (38/38 laboratory reference strains and 19/19 invasive clinical isolates) and 100 % exclusive; differentiating strains of S. pneumoniae species from other viridans group streptococci, including S. pseudopneumoniae. When applied to the analysis of a small number (n = 11) of clinical samples (blood culture positive for S. pneumoniae), the RPA assay was demonstrated to be both rapid and sensitive. The RPA assay developed in this work is shown to be as sensitive and as specific as PCR. In terms of reaction kinetics, the RPA assay is shown to exceed those of the PCR, with the RPA running to completion in 20 minutes and capable generating a positive signal in as little as 6 minutes. This work represents a potentially suitable assay for application in point-of-care settings.

Twitter Demographics

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Mendeley readers

The data shown below were compiled from readership statistics for 66 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United Kingdom 2 3%
Unknown 64 97%

Demographic breakdown

Readers by professional status Count As %
Researcher 14 21%
Student > Master 13 20%
Student > Ph. D. Student 11 17%
Student > Bachelor 7 11%
Student > Postgraduate 4 6%
Other 10 15%
Unknown 7 11%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 18 27%
Agricultural and Biological Sciences 12 18%
Medicine and Dentistry 7 11%
Immunology and Microbiology 5 8%
Nursing and Health Professions 3 5%
Other 15 23%
Unknown 6 9%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. 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 29 October 2015.
All research outputs
#13,818,651
of 18,008,158 outputs
Outputs from BMC Infectious Diseases
#4,075
of 6,357 outputs
Outputs of similar age
#191,326
of 292,866 outputs
Outputs of similar age from BMC Infectious Diseases
#385
of 584 outputs
Altmetric has tracked 18,008,158 research outputs across all sources so far. This one is in the 20th percentile – i.e., 20% of other outputs scored the same or lower than it.
So far Altmetric has tracked 6,357 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 7.7. This one is in the 28th percentile – i.e., 28% of its peers scored the same or lower than it.
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 292,866 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 29th percentile – i.e., 29% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 584 others from the same source and published within six weeks on either side of this one. This one is in the 24th percentile – i.e., 24% of its contemporaries scored the same or lower than it.