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. 2019 Dec;9(12):1462-1469.
doi: 10.1002/alr.22425. Epub 2019 Sep 4.

Detection and quantification of Staphylococcus in chronic rhinosinusitis

Brett Wagner Mackenzie  1 , Jesse Baker  1   2 , Richard G Douglas  1 , Michael W Taylor  2 , Kristi Biswas  2
Affiliations

Detection and quantification of Staphylococcus in chronic rhinosinusitis

Brett Wagner Mackenzie et al. Int Forum Allergy Rhinol. 2019 Dec.

Abstract

Background: The sinonasal microbiota has been implicated in chronic rhinosinusitis (CRS) pathogenesis, particularly related to the presence of Staphylococcus aureus. Staphylococcus epidermidis is also prevalent within the sinonasal microbiota and may inhibit S. aureus colonization. We investigated polymerase chain reaction (PCR) primer pairs for measuring absolute abundances of S. aureus and S. epidermidis, then compared bacterial community composition and absolute abundances of these species between CRS patients and controls.

Methods: Six candidate Staphylococcus species-specific primer pairs were tested in silico and in vitro against pure bacterial isolates. Quantitative PCR (qPCR) for absolute quantification of S. aureus, S. epidermidis, and overall bacterial load were assessed in 40 CRS (CRS without nasal polyposis [CRSsNP] = 22, CRS with nasal polyposis [CRSwNP] = 18) patients and 14 controls. Amplicon sequencing of the V3-V4 hypervariable regions of the 16S ribosomal RNA (rRNA) bacterial gene were conducted to investigate community composition.

Results: Primer pairs targeting the gmk gene of S. aureus and nrd gene from S. epidermidis were the most specific and sensitive primers. S. aureus (CRSsNP = 81.8% occurrence, CRSwNP = 83%, control = 92.9%) and S. epidermidis (CRSsNP = 95.5%, CRSwNP = 100%, control = 92.9%) were very prevalent, as indicated by qPCR results. Both CRSsNP and CRSwNP had significantly (p < 0.05) higher bacterial load when compared with controls (p < 0.05 for both). No significant correlation was observed between S. aureus and S. epidermidis abundances (p > 0.05).

Conclusion: Bacterial community sequencing detected Staphylococcus-assigned sequences in nearly all patients; however, it could not differentiate between S. aureus and S. epidermidis. Here, we present primer pairs that can distinguish between these species. We report a very high prevalence of S. aureus in both CRS patients and controls.

Keywords: Staphylococcus spp; bacteriology; chronic rhinosinusitis; paranasal sinuses; qPCR; statistics.

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References

    1. Grumann D, Nübel U, Bröker BM. Staphylococcus aureus toxins-their functions and genetics. Infect Genet Evol. 2014;21:583-592.
    1. Nakamura Y, Oscherwitz J, Cease KB, et al. Staphylococcus δ-toxin induces allergic skin disease by activating mast cells. Nature. 2013;503:397-401.
    1. Mulcahy ME, Geoghegan JA, Monk IR, et al. Nasal colonisation by Staphylococcus aureus depends upon clumping factor B binding to the squamous epithelial cell envelope protein loricrin. PLoS Pathog. 2012;8:e1003092.
    1. Peacock SJ, Moore CE, Justice A, et al. Virulent combinations of adhesin and toxin genes in natural populations of Staphylococcus aureus. Infect Immun. 2002;70:4987-4996.
    1. Burian M, Wolz C, Goerke C. Regulatory adaptation of Staphylococcus aureus during nasal colonization of humans. PLoS One. 2010;5:e10040.

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