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Development and validation of a real-time PCR assay for the detection of clinical acanthamoebae

Overview of attention for article published in BMC Research Notes, July 2017
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  • Above-average Attention Score compared to outputs of the same age and source (57th percentile)

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Title
Development and validation of a real-time PCR assay for the detection of clinical acanthamoebae
Published in
BMC Research Notes, July 2017
DOI 10.1186/s13104-017-2666-x
Pubmed ID
Authors

Nessika Karsenti, Rachel Lau, Andrew Purssell, Ann Chong-Kit, Marlou Cunanan, Jason Gasgas, Jinfang Tian, Amanda Wang, Filip Ralevski, Andrea K. Boggild

Abstract

Suboptimal agreement between molecular assays for the detection of Acanthamoeba spp. in clinical specimens has been demonstrated, and poor assay sensitivity directly imperils the vision of those affected by amoebic keratitis (AK) through delayed diagnosis. We sought to develop and validate a single Taqman real time PCR assay targeting the Acanthamoeba 18S rRNA gene that could be used to enhance sensitivity and specificity when paired with reference assays. Biobanked DNA from surplus delinked AK clinical specimens and 10 ATCC strains of Acanthamoeba was extracted. Sequence alignment of 66 18S rRNA regions from 12 species of Acanthamoeba known to cause keratitis informed design of a new TaqMan primer set. Performance of the new assay was compared to the 2 assays used currently in our laboratory. Among 24 Acanthamoeba-positive and 83 negative specimens by the CDC reference standard, performance characteristics of the newly designed primer set were as follows: sensitivity 100%, specificity 94%, PPV 82.8%, and NPV 100%. Compared to culture, sensitivity of the new primer set was 100%, and specificity 96%. No cross-reactivity of the primer set to non-acanthamoebae, including Balamuthia and Naegleria, was found. We have validated a real time PCR assay for the diagnosis of AK, and in doing so, have overcome important barriers to rapid and sensitive detection of acanthamoebae, including limited sensitivity and specificity of commonly used assays.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 40 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 40 100%

Demographic breakdown

Readers by professional status Count As %
Student > Master 8 20%
Researcher 5 13%
Student > Bachelor 4 10%
Professor > Associate Professor 3 8%
Student > Ph. D. Student 2 5%
Other 5 13%
Unknown 13 33%
Readers by discipline Count As %
Medicine and Dentistry 7 18%
Agricultural and Biological Sciences 5 13%
Engineering 5 13%
Biochemistry, Genetics and Molecular Biology 4 10%
Immunology and Microbiology 2 5%
Other 2 5%
Unknown 15 38%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 2. 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 10 August 2017.
All research outputs
#14,076,260
of 22,994,508 outputs
Outputs from BMC Research Notes
#1,871
of 4,284 outputs
Outputs of similar age
#169,903
of 316,684 outputs
Outputs of similar age from BMC Research Notes
#62
of 152 outputs
Altmetric has tracked 22,994,508 research outputs across all sources so far. This one is in the 37th percentile – i.e., 37% of other outputs scored the same or lower than it.
So far Altmetric has tracked 4,284 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.5. This one has gotten more attention than average, scoring higher than 54% 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 316,684 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 44th percentile – i.e., 44% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 152 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 57% of its contemporaries.