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A new real-time PCR protocol for detection of avian haemosporidians

Overview of attention for article published in Parasites & Vectors, July 2015
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About this Attention Score

  • Above-average Attention Score compared to outputs of the same age (55th percentile)
  • Above-average Attention Score compared to outputs of the same age and source (58th percentile)

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3 tweeters

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105 Mendeley
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Title
A new real-time PCR protocol for detection of avian haemosporidians
Published in
Parasites & Vectors, July 2015
DOI 10.1186/s13071-015-0993-0
Pubmed ID
Authors

Jeffrey A. Bell, Jason D. Weckstein, Alan Fecchio, Vasyl V. Tkach

Abstract

Birds possess the most diverse assemblage of haemosporidian parasites; including three genera, Plasmodium, Haemoproteus, and Leucocytozoon. Currently there are over 200 morphologically identified avian haemosporidian species, although true species richness is unknown due to great genetic diversity and insufficient sampling in highly diverse regions. Studies aimed at surveying haemosporidian diversity involve collecting and screening samples from hundreds to thousands of individuals. Currently, screening relies on microscopy and/or single or nested standard PCR. Although effective, these methods are time and resource consuming, and in the case of microscopy require substantial expertise. Here we report a newly developed real-time PCR protocol designed to quickly and reliably detect all three genera of avian haemosporidians in a single biochemical reaction. Using available DNA sequences from avian haemosporidians we designed primers R330F and R480RL, which flank a 182 base pair fragment of mitochondrial conserved rDNA. These primers were initially tested using real-time PCR on samples from Malawi, Africa, previously screened for avian haemosporidians using traditional nested PCR. Our real time protocol was further tested on 94 samples from the Cerrado biome of Brazil, previously screened using a single PCR assay for haemosporidian parasites. These samples were also amplified using modified nested PCR protocols, allowing for comparisons between the three different screening methods (single PCR, nested PCR, real-time PCR). The real-time PCR protocol successfully identified all three genera of avian haemosporidians from both single and mixed infections previously detected from Malawi. There was no significant difference between the three different screening protocols used for the 94 samples from the Brazilian Cerrado (χ (2) = 0.3429, df = 2, P = 0.842). After proving effective, the real-time protocol was used to screen 2113 Brazilian samples, identifying 693 positive samples. Our real-time PCR assay proved as effective as two widely used molecular screening techniques, single PCR and nested PCR. However, the real-time protocol has the distinct advantage of detecting all three genera in a single reaction, which significantly increases efficiency by greatly decreasing screening time and cost. Our real-time PCR protocol is therefore a valuable tool in the quickly expanding field of avian haemosporidian research.

Twitter Demographics

The data shown below were collected from the profiles of 3 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 105 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Lithuania 2 2%
United States 2 2%
New Zealand 1 <1%
Unknown 100 95%

Demographic breakdown

Readers by professional status Count As %
Student > Master 21 20%
Researcher 21 20%
Student > Ph. D. Student 19 18%
Student > Bachelor 16 15%
Student > Doctoral Student 10 10%
Other 13 12%
Unknown 5 5%
Readers by discipline Count As %
Agricultural and Biological Sciences 61 58%
Veterinary Science and Veterinary Medicine 9 9%
Environmental Science 9 9%
Biochemistry, Genetics and Molecular Biology 9 9%
Immunology and Microbiology 3 3%
Other 6 6%
Unknown 8 8%

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 19 July 2015.
All research outputs
#2,391,634
of 5,372,803 outputs
Outputs from Parasites & Vectors
#614
of 1,676 outputs
Outputs of similar age
#81,146
of 188,978 outputs
Outputs of similar age from Parasites & Vectors
#38
of 107 outputs
Altmetric has tracked 5,372,803 research outputs across all sources so far. This one has received more attention than most of these and is in the 54th percentile.
So far Altmetric has tracked 1,676 research outputs from this source. They receive a mean Attention Score of 3.3. This one has gotten more attention than average, scoring higher than 60% 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 188,978 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 55% of its contemporaries.
We're also able to compare this research output to 107 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 58% of its contemporaries.