Kramná Lenka, Dřevínek Pavel, Lin Jake, Kulich Michal, Cinek Ondrej
Department of Paediatrics, 2nd Faculty of Medicine, Charles University in Prague and University Hospital Motol, V Úvalu 84, 15006, Prague 5, Czech Republic.
Department of Medical Microbiology, 2nd Faculty of Medicine, Charles University in Prague and University Hospital Motol, Prague, Czech Republic.
Folia Microbiol (Praha). 2018 Mar;63(2):237-248. doi: 10.1007/s12223-017-0562-3. Epub 2017 Nov 10.
The lung in cystic fibrosis (CF) is home to numerous pathogens that shorten the lives of patients. The aim of the present study was to assess changes in the lung bacteriome following antibiotic therapy targeting Pseudomonas aeruginosa in children with CF. The study included nine children (9-18 years) with CF who were treated for their chronic or intermittent positivity for Pseudomonas aeruginosa. The bacteriomes were determined in 16 pairs of sputa collected at the beginning and at the end of a course of intravenous antibiotic therapy via deep sequencing of the variable region 4 of the 16S rRNA gene, and the total bacterial load and selected specific pathogens were assessed using quantitative real-time PCR. The effect of antipseudomonal antibiotics was observable as a profound decrease in the total 16S rDNA load (p = 0.001) as well as in a broad range of individual taxa including Staphylococcus aureus (p = 0.03) and several members of the Streptococcus mitis group (S. oralis, S. mitis, and S. infantis) (p = 0.003). Improvements in forced expiratory volume (FEV) were associated with an increase in Granulicatella sp. (p = 0.004), whereas a negative association was noted between the total bacterial load and white blood cell count (p = 0.007). In conclusion, the data show how microbial communities differ in reaction to antipseudomonal treatment, suggesting that certain rare species may be associated with clinical parameters. Our work also demonstrates the utility of absolute quantification of bacterial load in addition to the 16S rDNA profiling.
囊性纤维化(CF)患者的肺部存在众多病原体,这些病原体缩短了患者的寿命。本研究的目的是评估针对CF患儿铜绿假单胞菌的抗生素治疗后肺部细菌群落的变化。该研究纳入了9名年龄在9至18岁之间的CF患儿,他们因慢性或间歇性铜绿假单胞菌阳性而接受治疗。通过对16S rRNA基因可变区4进行深度测序,测定了静脉抗生素治疗疗程开始和结束时采集的16对痰液中的细菌群落,并使用定量实时PCR评估了总细菌载量和选定的特定病原体。抗铜绿假单胞菌抗生素的效果表现为16S rDNA总载量显著下降(p = 0.001),以及包括金黄色葡萄球菌(p = 0.03)和缓症链球菌组的几个成员(口腔链球菌、缓症链球菌和婴儿链球菌)(p = 0.003)在内的多种个体分类群减少。用力呼气量(FEV)的改善与Granulicatella菌属增加相关(p = 0.004),而总细菌载量与白细胞计数之间存在负相关(p = 0.007)。总之,数据显示了微生物群落在对抗铜绿假单胞菌治疗的反应上存在差异,表明某些罕见物种可能与临床参数相关。我们的工作还证明了除16S rDNA分析外,细菌载量绝对定量的实用性。