西班牙儿童伴自发性耳漏的急性中耳炎的细菌病原学：一项多中心研究的分子学见解。

文献信息

中文摘要

未标注： 急性中耳炎（AOM）是儿童期最常见的感染性疾病之一，肺炎链球菌和流感嗜血杆菌是最常涉及的病原体。在2019冠状病毒病（COVID-19）大流行以及13价肺炎球菌结合疫苗（PCV13）纳入国家免疫规划之后，已有关于肺炎球菌侵袭性疾病变化的描述，但关于AOM的数据仍然有限。本研究旨在评估当前儿童AOM的细菌病原学。从就诊于巴塞罗那南部13个初级保健中心的AOM患儿中采集自发性耳漏液（SOF）样本和可选的鼻咽样本（NP）。采用常规培养和PCR进行病原体检测。对最常见的病原体（化脓性链球菌、肺炎链球菌和流感嗜血杆菌）进行了抗菌药物敏感性试验和全基因组测序。共获得145份SOF和83份NP样本。SOF样本中通过培养检测到细菌生长者占80.7%，通过分子方法检测到者占91.7%。主要病原体为流感嗜血杆菌（40.7%）、化脓性链球菌（35.2%）和肺炎链球菌（33.8%）。血清型3（主要为GPSC12-CC180）是最常见的肺炎球菌血清型（15.6%），尽管疫苗接种覆盖率很高，但已鉴定血清型中有23.8%包含在PCV13中。所分析的化脓性链球菌大多数（43.1%）属于emm1-ST28基因型，所分析的流感嗜血杆菌分离株中94.1%为不可分型。这些发现表明在PCV13后时代AOM病原学发生了显著转变，流感嗜血杆菌成为主要病原体，且化脓性链球菌AOM增加。持续监测对于为未来的疫苗接种和预防策略提供信息至关重要。
重要性： 这项多中心研究为接种13价肺炎球菌结合疫苗的儿童中急性中耳炎（AOM）的当前流行病学提供了新见解。首先，它强调了分子方法在病原体检测中的作用。其次，它增加了鼻咽样本与自发性耳漏液样本之间缺乏一致性（尤其是对肺炎链球菌）的新证据。第三，全基因组分析证实GSPC12-clade Ia占优势，是血清型3肺炎球菌所致疫苗失败的主要贡献者。第四，我们描述了与近期AOM中化脓性链球菌激增相关的谱系，这些谱系不同于本地区引起侵袭性疾病的谱系。最后，我们的发现证实了不可分型流感嗜血杆菌目前作为接种疫苗儿童中AOM首要病因的优势地位。

英文摘要

UNLABELLED: Acute otitis media (AOM) is one of the most prevalent infectious diseases in childhood, with Streptococcus pneumoniae and Haemophilus influenzae being the most commonly involved pathogens. Following the coronavirus disease 2019 (COVID-19) pandemic and the incorporation of 13-valent pneumococcal conjugate vaccine (PCV13) into the national immunization program, changes in pneumococcal invasive disease have been described, but data on AOM remain limited. This study aimed to evaluate the current bacterial etiology of pediatric AOM. Spontaneous otorrhea fluid (SOF) samples and optional nasopharyngeal samples (NP) were collected from children with AOM attending 13 primary care centers in southern Barcelona. Conventional culture and PCR were used for pathogen detection. Most frequent pathogens (Streptococcus pyogenes, S. pneumoniae, and H. influenzae) underwent antimicrobial susceptibility testing and whole-genome sequencing. A total of 145 SOF and 83 NP samples were obtained. Bacterial growth in SOF samples was detected by culture in 80.7% of the samples and by molecular methods in 91.7%. The predominant pathogens were H. influenzae (40.7%), S. pyogenes (35.2%), and S. pneumoniae (33.8%). Serotype 3 (mainly GPSC12-CC180) was the most frequent pneumococcal serotype (15.6%), and 23.8% of identified serotypes were included in PCV13 despite high vaccination coverage. Most of the S. pyogenes analyzed (43.1%) belonged to the emm1-ST28 genotype, and 94.1% of H. influenzae isolates analyzed were non-typeable. These findings demonstrate a marked shift in AOM etiology in the post-PCV13 era, with H. influenzae emerging as the leading pathogen and an increase in S. pyogenes AOM. Continued surveillance is essential to inform future vaccination and prevention strategies.
IMPORTANCE: This multicenter study provides new insights into the current epidemiology of acute otitis media (AOM) in children vaccinated with the 13-valent pneumococcal conjugate vaccine. First, it underscores the role of molecular methods in pathogen detection. Second, it adds new evidence of the lack of concordance between nasopharyngeal and spontaneous otorrhea fluid samples, particularly for S. pneumoniae. Third, whole-genome analysis confirmed the predominance of the GSPC12-clade Ia as the major contributor to the vaccine failures due to serotype 3 pneumococci. Fourth, we characterized the S. pyogenes lineages associated with its recent upsurge in AOM, which differs from those causing invasive disease in our area. Finally, our findings confirm the current dominance of non-typeable H. influenzae as the leading cause of AOM in vaccinated children.