Department of Surgery, The University of Auckland, Auckland, New Zealand.
School of Biological Sciences and Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Auckland, New Zealand.
Int Forum Allergy Rhinol. 2019 Dec;9(12):1462-1469. doi: 10.1002/alr.22425. Epub 2019 Sep 4.
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.
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.
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).
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.
鼻旁窦微生物群与慢性鼻旁窦炎（CRS）的发病机制有关，特别是与金黄色葡萄球菌的存在有关。表皮葡萄球菌也在鼻旁窦微生物群中普遍存在，并可能抑制金黄色葡萄球菌定植。我们研究了用于测量金黄色葡萄球菌和表皮葡萄球菌绝对丰度的聚合酶链反应（PCR）引物对，然后比较了 CRS 患者和对照组之间的细菌群落组成和这些物种的绝对丰度。
在体外和体外测试了 6 对候选葡萄球菌种特异性引物对，针对纯细菌分离株。在 40 例 CRS（无鼻息肉的 CRS [CRSsNP] = 22 例，有鼻息肉的 CRS [CRSwNP] = 18 例）患者和 14 例对照中评估了金黄色葡萄球菌、表皮葡萄球菌和总细菌负荷的绝对定量 qPCR（qPCR）。进行 16S 核糖体 RNA（rRNA）细菌基因的 V3-V4 高变区扩增子测序，以研究群落组成。
针对金黄色葡萄球菌的 gmk 基因和表皮葡萄球菌的 nrd 基因的引物对是最特异和敏感的引物。qPCR 结果表明，金黄色葡萄球菌（CRSsNP = 81.8%发生，CRSwNP = 83%，对照 = 92.9%）和表皮葡萄球菌（CRSsNP = 95.5%，CRSwNP = 100%，对照 = 92.9%）非常普遍。与对照组相比，CRSsNP 和 CRSwNP 的细菌负荷均显著升高（均 p < 0.05）。金黄色葡萄球菌和表皮葡萄球菌丰度之间未观察到显著相关性（均 p > 0.05）。
细菌群落测序在几乎所有患者中均检测到葡萄球菌分配序列；然而，它不能区分金黄色葡萄球菌和表皮葡萄球菌。在这里，我们提出了可以区分这些物种的引物对。我们报告了金黄色葡萄球菌在 CRS 患者和对照组中均非常普遍。
