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Detection of viable Plasmodium ookinetes in the midguts of Anopheles coluzzi using PMA-qrtPCR

Overview of attention for article published in Parasites & Vectors, September 2015
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  • Good Attention Score compared to outputs of the same age (70th percentile)

Mentioned by

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6 tweeters
peer_reviews
1 peer review site

Citations

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5 Dimensions

Readers on

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43 Mendeley
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Title
Detection of viable Plasmodium ookinetes in the midguts of Anopheles coluzzi using PMA-qrtPCR
Published in
Parasites & Vectors, September 2015
DOI 10.1186/s13071-015-1087-8
Pubmed ID
Authors

Tibebu Habtewold, Zoe Groom, Luc Duchateau, George K. Christophides

Abstract

Mosquito infection with malaria parasites depends on complex interactions between the mosquito immune response, the parasite developmental program and the midgut microbiota. Simultaneous monitoring of the parasite and bacterial dynamics is important when studying these interactions. PCR based methods of genomic DNA (gDNA) have been widely used, but their inability to discriminate between live and dead cells compromises their application. The alternative method of quantification of mRNA mainly reports on cell activity rather than density. Quantitative real-time (qrt) PCR in combination with Propidium Monoazide (PMA) treatment (PMA-qrtPCR) has been previously used for selectively enumerating viable microbial cells. PMA penetrates damaged cell membranes and intercalates in the DNA inhibiting its PCR amplification. Here, we tested the potential of PMA-qrtPCR to discriminate between and quantify live and dead Plasmodium berghei malarial parasites and commensal bacteria in the midgut of Anopheles coluzzii Coetzee & Wilkerson 2013 (formerly An. gambiae M-form). By combining microscopic observations with reverse transcriptase PCR (RT-PCR) we reveal that, in addition to gDNA, mRNA from dead parasites also persists inside the mosquito midgut, therefore its quantification cannot accurately reflect live-only parasites at the time of monitoring. In contrast, pre-treating the samples with PMA selectively inhibited qrtPCR amplification of parasite gDNA, with about 15 cycles (Ct-value) difference between PMA-treated and control samples. The limit of detection corresponds to 10 Plasmodium ookinetes. Finally, we show that the PMA-qrtPCR method can be used to quantify bacteria that are present in the mosquito midgut. The PMA-qrtPCR is a suitable method for quantification of viable parasites and bacteria in the midgut of Anopheles mosquitoes. The method will be valuable when studying the molecular interactions between the mosquito, the malaria parasite and midgut microbiota.

Twitter Demographics

The data shown below were collected from the profiles of 6 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 43 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Spain 1 2%
Ghana 1 2%
Unknown 41 95%

Demographic breakdown

Readers by professional status Count As %
Researcher 9 21%
Student > Ph. D. Student 9 21%
Student > Master 8 19%
Student > Postgraduate 4 9%
Student > Bachelor 3 7%
Other 5 12%
Unknown 5 12%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 11 26%
Agricultural and Biological Sciences 10 23%
Medicine and Dentistry 7 16%
Immunology and Microbiology 4 9%
Pharmacology, Toxicology and Pharmaceutical Science 1 2%
Other 4 9%
Unknown 6 14%

Attention Score in Context

This research output has an Altmetric Attention Score of 4. 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 08 February 2016.
All research outputs
#4,874,875
of 16,518,952 outputs
Outputs from Parasites & Vectors
#1,144
of 4,395 outputs
Outputs of similar age
#72,412
of 249,303 outputs
Outputs of similar age from Parasites & Vectors
#1
of 1 outputs
Altmetric has tracked 16,518,952 research outputs across all sources so far. This one has received more attention than most of these and is in the 70th percentile.
So far Altmetric has tracked 4,395 research outputs from this source. They receive a mean Attention Score of 4.8. This one has gotten more attention than average, scoring higher than 73% 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 249,303 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 70% of its contemporaries.
We're also able to compare this research output to 1 others from the same source and published within six weeks on either side of this one. This one has scored higher than all of them