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 journalArticleResearchpeer-review

    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 languageEnglish
    Pages (from-to)585-593
    Number of pages9
    JournalFuture Microbiology
    Volume12
    Issue number7
    DOIs
    Publication statusPublished - Jun 2017

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    Haemophilus
    Haemophilus influenzae
    Real-Time Polymerase Chain Reaction
    Polymerase Chain Reaction
    Fucose
    Multiplex Polymerase Chain Reaction
    Genome

    Cite this

    Price, E. P., Harris, T. M., Spargo, J., Nosworthy, E., Beissbarth, J., Chang, A. B., ... Sarovich, D. S. (2017). Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR. Future Microbiology, 12(7), 585-593. https://doi.org/10.2217/fmb-2016-0215
    Price, Erin P. ; Harris, Tegan M. ; Spargo, Jessie ; Nosworthy, Elizabeth ; Beissbarth, Jemima ; Chang, Anne B. ; Smith-Vaughan, Heidi C. ; Sarovich, Derek S. / Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR. In: Future Microbiology. 2017 ; Vol. 12, No. 7. pp. 585-593.
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    Simultaneous identification of Haemophilus influenzae and Haemophilus haemolyticus using real-time PCR. / Price, Erin P.; Harris, Tegan M.; Spargo, Jessie; Nosworthy, Elizabeth; Beissbarth, Jemima; Chang, Anne B.; Smith-Vaughan, Heidi C.; Sarovich, Derek S.

    In: Future Microbiology, Vol. 12, No. 7, 06.2017, p. 585-593.

    Research output: Contribution to journalArticleResearchpeer-review

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    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.

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    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.

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