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Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples

Overview of attention for article published in Brazilian Journal of Microbiology, July 2018
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Title
Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples
Published in
Brazilian Journal of Microbiology, July 2018
DOI 10.1016/j.bjm.2017.09.004
Pubmed ID
Authors

Bruno Alonso Miotto, Aline Santana da Hora, Sueli Akemi Taniwaki, Paulo Eduardo Brandão, Marcos Bryan Heinemann, Mitika Kuribayashi Hagiwara

Abstract

A modified TaqMan real-time polymerase chain reaction targeting a 138bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleotide sequencing analysis. The overall agreement between the assays was 94.8% (confidence interval [CI] 95% 88-100%). The newly developed assay presented 91.6% (CI 95% 71.5-98.5%) relative sensitivity (22[+] lipl32 PCR/24[+] 16S RNA and sequencing), 100% (CI 95% 96.3-100%) relative specificity and 98.7% accuracy (CI 95% 94.8-100%). The lipl32 assay was able to detect and quantify at least 10 genome equivalents/reaction. DNA extracted from 17 pathogenic Leptospira spp., 8 intermediate/saprophytic strains and 21 different pathogenic microorganisms were also tested using the lipl32 assay, resulting in amplification exclusively for pathogenic leptospiral strains. The results also demonstrated high intra and inter-assay reproducibility (coefficient of variation 1.50 and 1.12, respectively), thereby qualifying the newly developed assay as a highly sensitive, specific and reliable diagnostic tool for leptospiral infection in dogs using urine specimens.

Twitter Demographics

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Mendeley readers

The data shown below were compiled from readership statistics for 58 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 58 100%

Demographic breakdown

Readers by professional status Count As %
Student > Bachelor 10 17%
Student > Master 8 14%
Student > Ph. D. Student 6 10%
Student > Postgraduate 4 7%
Researcher 3 5%
Other 8 14%
Unknown 19 33%
Readers by discipline Count As %
Veterinary Science and Veterinary Medicine 20 34%
Agricultural and Biological Sciences 6 10%
Biochemistry, Genetics and Molecular Biology 4 7%
Medicine and Dentistry 3 5%
Environmental Science 1 2%
Other 5 9%
Unknown 19 33%

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 19 October 2018.
All research outputs
#10,868,832
of 13,644,952 outputs
Outputs from Brazilian Journal of Microbiology
#284
of 441 outputs
Outputs of similar age
#283,502
of 393,189 outputs
Outputs of similar age from Brazilian Journal of Microbiology
#10
of 19 outputs
Altmetric has tracked 13,644,952 research outputs across all sources so far. This one is in the 11th percentile – i.e., 11% of other outputs scored the same or lower than it.
So far Altmetric has tracked 441 research outputs from this source. They receive a mean Attention Score of 2.2. This one is in the 21st percentile – i.e., 21% of its peers scored the same or lower than it.
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We're also able to compare this research output to 19 others from the same source and published within six weeks on either side of this one. This one is in the 10th percentile – i.e., 10% of its contemporaries scored the same or lower than it.