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

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Price, Erin P
Harris, Tegan M
Spargo, Jessie C
Nosworthy, Elizabeth
Beissbarth, Jemima
Chang, Anne B
Smith-Vaughan, Heidi C
Sarovich, Derek S
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2017
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Abstract

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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Future Microbiology

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12

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7

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© The Author(s) 2017. This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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Microbiology

Medical microbiology

Science & Technology

Life Sciences & Biomedicine

fucP

genotyping

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Price, EP; Harris, TM; Spargo, JC; Nosworthy, E; Beissbarth, J; Chang, AB; Smith-Vaughan, HC; Sarovich, DS, Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR, Future Microbiology, 2017, 12 (7), pp. 585-593

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