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SYBR Green-Based Real-Time Reverse Transcription-PCR for Typing and Subtyping of All Hemagglutinin and Neuraminidase Genes of Avian Influenza Viruses and Comparison to Standard Serological Subtyping…

Overview of attention for article published in Journal of Clinical Microbiology, October 2011
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  • In the top 25% of all research outputs scored by Altmetric
  • High Attention Score compared to outputs of the same age (80th percentile)
  • Good Attention Score compared to outputs of the same age and source (69th percentile)

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24 Facebook pages

Citations

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20 Dimensions

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32 Mendeley
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Title
SYBR Green-Based Real-Time Reverse Transcription-PCR for Typing and Subtyping of All Hemagglutinin and Neuraminidase Genes of Avian Influenza Viruses and Comparison to Standard Serological Subtyping Tests
Published in
Journal of Clinical Microbiology, October 2011
DOI 10.1128/jcm.01195-11
Pubmed ID
Authors

Kenji Tsukamoto, Panei Carlos Javier, Makiko Shishido, Daigo Noguchi, John Pearce, Hyun-Mi Kang, Ok Mi Jeong, Youn-Jeong Lee, Koji Nakanishi, Takayoshi Ashizawa

Abstract

Continuing outbreaks of H5N1 highly pathogenic (HP) avian influenza virus (AIV) infections of wild birds and poultry worldwide emphasize the need for global surveillance of wild birds. To support the future surveillance activities, we developed a SYBR green-based, real-time reverse transcriptase PCR (rRT-PCR) for detecting nucleoprotein (NP) genes and subtyping 16 hemagglutinin (HA) and 9 neuraminidase (NA) genes simultaneously. Primers were improved by focusing on Eurasian or North American lineage genes; the number of mixed-base positions per primer was set to five or fewer, and the concentration of each primer set was optimized empirically. Also, 30 cycles of amplification of 1:10 dilutions of cDNAs from cultured viruses effectively reduced minor cross- or nonspecific reactions. Under these conditions, 346 HA and 345 NA genes of 349 AIVs were detected, with average sensitivities of NP, HA, and NA genes of 10(1.5), 10(2.3), and 10(3.1) 50% egg infective doses, respectively. Utility of rRT-PCR for subtyping AIVs was compared with that of current standard serological tests by using 104 recent migratory duck virus isolates. As a result, all HA genes and 99% of the NA genes were genetically subtyped, while only 45% of HA genes and 74% of NA genes were serologically subtyped. Additionally, direct subtyping of AIVs in fecal samples was possible by 40 cycles of amplification: approximately 70% of HA and NA genes of NP gene-positive samples were successfully subtyped. This validation study indicates that rRT-PCR with optimized primers and reaction conditions is a powerful tool for subtyping varied AIVs in clinical and cultured samples.

Mendeley readers

The data shown below were compiled from readership statistics for 32 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Uzbekistan 1 3%
United States 1 3%
Denmark 1 3%
Unknown 29 91%

Demographic breakdown

Readers by professional status Count As %
Researcher 11 34%
Student > Master 5 16%
Student > Bachelor 3 9%
Professor 2 6%
Other 2 6%
Other 6 19%
Unknown 3 9%
Readers by discipline Count As %
Agricultural and Biological Sciences 13 41%
Veterinary Science and Veterinary Medicine 5 16%
Biochemistry, Genetics and Molecular Biology 3 9%
Medicine and Dentistry 3 9%
Immunology and Microbiology 2 6%
Other 4 13%
Unknown 2 6%

Attention Score in Context

This research output has an Altmetric Attention Score of 6. 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 30 July 2012.
All research outputs
#2,146,095
of 12,360,070 outputs
Outputs from Journal of Clinical Microbiology
#1,224
of 7,013 outputs
Outputs of similar age
#21,274
of 122,369 outputs
Outputs of similar age from Journal of Clinical Microbiology
#13
of 63 outputs
Altmetric has tracked 12,360,070 research outputs across all sources so far. Compared to these this one has done well and is in the 78th percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 7,013 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.6. This one has gotten more attention than average, scoring higher than 74% 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 122,369 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 80% of its contemporaries.
We're also able to compare this research output to 63 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 69% of its contemporaries.