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 journalArticle

    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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