Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR

Erin P Price; T M Harris; J Spargo; E Nosworthy; J Beizzbarth; A B Chang; H C Smith-Vaughan; Derek S Sarovich

doi:10.2217/fmb-2016-0215

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Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR

Journal article

Open access

Peer reviewed

Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR

Erin P Price, T M Harris, J Spargo, E Nosworthy, J Beizzbarth, A B Chang, H C Smith-Vaughan and Derek S Sarovich

Future Microbiology, Vol.12(7), pp.585-593

2017

DOI: https://doi.org/10.2217/fmb-2016-0215

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Abstract

fucP

genotyping

Haemophilus haemolyticus

Haemophilus influenzae

hypD

multiplex

real-time PCR

siaT

TaqMan

triplex

Aim: To design a highly specific and sensitive multiplex real-time PCR assay for the differentiation of the pathogen Haemophilus influenzae from its nonpathogenic near-neighbor Haemophilus haemolyticus. Materials & methods: A comparison of 380 Haemophilus spp. genomes was used to identify loci specific for each species. Novel PCR assays targeting H. haemolyticus (hypD) and H. influenzae (siaT) were designed. Results & discussion: PCR screening across 143 isolates demonstrated 100% specificity for hypD and siaT. These two assays were multiplexed with the recently described fucP assay for further differentiation among H. influenzae. Conclusion: The triplex assay provides rapid, unambiguous, sensitive and highly specific genotyping results for the simultaneous detection of hypD and siaT, including fucose-positive H. influenzae (fucP), in a single PCR.

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Title: Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR
Authors: Erin P Price (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
T M Harris (Author) - Menzies School of Health Research
J Spargo (Author) - Menzies School of Health Research
E Nosworthy (Author) - Menzies School of Health Research
J Beizzbarth (Author) - Menzies School of Health Research
A B Chang (Author) - Menzies School of Health Research
H C Smith-Vaughan (Author) - Menzies School of Health Research
Derek S Sarovich (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Publication details: Future Microbiology, Vol.12(7), pp.585-593
Publisher: Future Medicine Ltd.
Date published: 2017
DOI: 10.2217/fmb-2016-0215
ISSN: 1746-0913
Copyright note: Copyright © 2017 Price, Harris, Spargo, Nosworthy, Beissbarth, Chang, Smith-Vaughan & Sarovich. This work is licensed under the Attribution-NonCommercialNoDerivatives 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99451281302621
Output Type: Journal article

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