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

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

doi:10.2217/fmb-2016-0215

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

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

Research output: Contribution to journal › Article › peer-review

30 Citations (Scopus)

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.

Original language	English
Pages (from-to)	585-593
Number of pages	9
Journal	Future Microbiology
Volume	12
Issue number	7
DOIs	https://doi.org/10.2217/fmb-2016-0215
Publication status	Published - Jun 2017

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title = "Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR",

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.",

keywords = "fucP, genotyping, Haemophilus haemolyticus, Haemophilus influenzae, hypD, multiplex, real-time PCR, siaT, TaqMan, triplex",

author = "Price, {Erin P.} and Harris, {Tegan M.} and Jessie Spargo and Elizabeth Nosworthy and Jemima Beissbarth and Chang, {Anne B.} and Smith-Vaughan, {Heidi C.} and Sarovich, {Derek S.}",

year = "2017",

month = jun,

doi = "10.2217/fmb-2016-0215",

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pages = "585--593",

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T1 - Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR

AU - Price, Erin P.

AU - Harris, Tegan M.

AU - Spargo, Jessie

AU - Nosworthy, Elizabeth

AU - Beissbarth, Jemima

AU - Chang, Anne B.

AU - Smith-Vaughan, Heidi C.

AU - Sarovich, Derek S.

PY - 2017/6

Y1 - 2017/6

N2 - 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.

AB - 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.

KW - fucP

KW - genotyping

KW - Haemophilus haemolyticus

KW - Haemophilus influenzae

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KW - TaqMan

KW - triplex

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JO - Future Microbiology

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ER -

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

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