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Internal control for real-time polymerase chain reaction based on MS2 bacteriophage for RNA viruses diagnostics

Overview of attention for article published in Memórias do Instituto Oswaldo Cruz, April 2017
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  • Above-average Attention Score compared to outputs of the same age and source (58th percentile)

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2 tweeters

Citations

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56 Mendeley
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Title
Internal control for real-time polymerase chain reaction based on MS2 bacteriophage for RNA viruses diagnostics
Published in
Memórias do Instituto Oswaldo Cruz, April 2017
DOI 10.1590/0074-02760160380
Pubmed ID
Authors

Miriam Ribas Zambenedetti, Daniela Parada Pavoni, Andreia Cristine Dallabona, Alejandro Correa Dominguez, Celina de Oliveira Poersch, Stenio Perdigão Fragoso, Marco Aurélio Krieger

Abstract

Real-time reverse transcription polymerase chain reaction (RT-PCR) is routinely used to detect viral infections. In Brazil, it is mandatory the use of nucleic acid tests to detect hepatitis C virus (HCV), hepatitis B virus and human immunodeficiency virus in blood banks because of the immunological window. The use of an internal control (IC) is necessary to differentiate the true negative results from those consequent from a failure in some step of the nucleic acid test. The aim of this study was the construction of virus-modified particles, based on MS2 bacteriophage, to be used as IC for the diagnosis of RNA viruses. The MS2 genome was cloned into the pET47b(+) plasmid, generating pET47b(+)-MS2. MS2-like particles were produced through the synthesis of MS2 RNA genome by T7 RNA polymerase. These particles were used as non-competitive IC in assays for RNA virus diagnostics. In addition, a competitive control for HCV diagnosis was developed by cloning a mutated HCV sequence into the MS2 replicase gene of pET47b(+)-MS2, which produces a non-propagating MS2 particle. The utility of MS2-like particles as IC was evaluated in a one-step format multiplex real-time RT-PCR for HCV detection. We demonstrated that both competitive and non-competitive IC could be successfully used to monitor the HCV amplification performance, including the extraction, reverse transcription, amplification and detection steps, without compromising the detection of samples with low target concentrations. In conclusion, MS2-like particles generated by this strategy proved to be useful IC for RNA virus diagnosis, with advantage that they are produced by a low cost protocol. An attractive feature of this system is that it allows the construction of a multicontrol by the insertion of sequences from more than one pathogen, increasing its applicability for diagnosing different RNA viruses.

Twitter Demographics

The data shown below were collected from the profiles of 2 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 56 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Brazil 1 2%
Unknown 55 98%

Demographic breakdown

Readers by professional status Count As %
Researcher 15 27%
Student > Bachelor 9 16%
Student > Ph. D. Student 6 11%
Student > Master 5 9%
Other 4 7%
Other 10 18%
Unknown 7 13%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 11 20%
Agricultural and Biological Sciences 8 14%
Immunology and Microbiology 6 11%
Business, Management and Accounting 4 7%
Social Sciences 3 5%
Other 13 23%
Unknown 11 20%

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 25 August 2017.
All research outputs
#7,850,606
of 12,511,869 outputs
Outputs from Memórias do Instituto Oswaldo Cruz
#599
of 862 outputs
Outputs of similar age
#147,725
of 258,557 outputs
Outputs of similar age from Memórias do Instituto Oswaldo Cruz
#5
of 12 outputs
Altmetric has tracked 12,511,869 research outputs across all sources so far. This one is in the 23rd percentile – i.e., 23% of other outputs scored the same or lower than it.
So far Altmetric has tracked 862 research outputs from this source. They receive a mean Attention Score of 3.2. This one is in the 30th percentile – i.e., 30% 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 258,557 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 33rd percentile – i.e., 33% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 12 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.