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    Comparison of established diagnostic methodologies and a novel bacterialsmpbreal-time pcr assay for specific detection of haemophilus influenzae isolates associated with respiratory tract infections

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    Date
    2015-06-24
    Author
    Reddington, Kate
    Schwenk, Stefan
    Tuite, Nina
    Platt, Gareth
    Davar, Danesh
    Coughlan, Helena
    Personne, Yoann
    Gant, Vanya
    Enne, Virve I.
    Zumla, Alimuddin
    Barry, Thomas
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    Cited 3 times in Scopus (view citations)
    
    Recommended Citation
    Reddington, Kate; Schwenk, Stefan; Tuite, Nina; Platt, Gareth; Davar, Danesh; Coughlan, Helena; Personne, Yoann; Gant, Vanya; Enne, Virve I. Zumla, Alimuddin; Barry, Thomas (2015). Comparison of established diagnostic methodologies and a novel bacterialsmpbreal-time pcr assay for specific detection of haemophilus influenzae isolates associated with respiratory tract infections. Journal of Clinical Microbiology 53 (9), 2854-2860
    Published Version
    https://jcm.asm.org/content/53/9/2854.full.pdf
    Abstract
    Haemophilus influenzae is a significant causative agent of respiratory tract infections (RTI) worldwide. The development of a rapid H. influenzae diagnostic assay that would allow for the implementation of infection control measures and also improve antimicrobial stewardship for patients is required. A number of nucleic acid diagnostics approaches that detect H. influenzae in RTIs have been described in the literature; however, there are reported specificity and sensitivity limitations for these assays. In this study, a novel real-time PCR diagnostic assay targeting the smpB gene was designed to detect all serogroups of H. influenzae. The assay was validated using a panel of well-characterized Haemophilus spp. Subsequently, 44 Haemophilus clinical isolates were collected, and 36 isolates were identified as H. influenzae using a gold standard methodology that combined the results of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and a fucK diagnostic assay. Using the novel smpB diagnostic assay, 100% concordance was observed with the gold standard, demonstrating a sensitivity of 100% (95% confidence interval [CI], 90.26% to 100.00%) and a specificity of 100% (95% CI, 63.06% to 100.00%) when used on clinical isolates. To demonstrate the clinical utility of the diagnostic assay presented, a panel of lower RTI samples (n = 98) were blindly tested with the gold standard and smpB diagnostic assays. The results generated were concordant for 94/98 samples tested, demonstrating a sensitivity of 90.91% (95% CI, 78.33% to 97.47%) and a specificity of 100% (95% CI, 93.40% to 100.00%) for the novel smpB assay when used directly on respiratory specimens.
    URI
    http://hdl.handle.net/10379/13613
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