Premium
Detection and quantification of Staphylococcus in chronic rhinosinusitis
Author(s) -
Wagner Mackenzie Brett,
Baker Jesse,
Douglas Richard G.,
Taylor Michael W.,
Biswas Kristi
Publication year - 2019
Publication title -
international forum of allergy and rhinology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.503
H-Index - 46
eISSN - 2042-6984
pISSN - 2042-6976
DOI - 10.1002/alr.22425
Subject(s) - staphylococcus epidermidis , staphylococcus aureus , microbiology and biotechnology , amplicon , medicine , hypervariable region , chronic rhinosinusitis , polymerase chain reaction , staphylococcus , staphylococcal infections , primer (cosmetics) , pathogenesis , biology , immunology , gene , bacteria , genetics , chemistry , organic chemistry , antibody
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.