DDL Diagnostic Laboratory, 2288 ER, Rijswijk, The Netherlands.
Department of Medical Microbiology, Haaglanden Medical Centre Bronovo, 2597 AX, The Hague, The Netherlands.
Eur J Clin Microbiol Infect Dis. 2019 Apr;38(4):747-754. doi: 10.1007/s10096-019-03511-4. Epub 2019 Feb 20.
In clinical practice, the diagnosis of lower respiratory tract infections (LRTIs) is based on culture. The aim of this study was to evaluate whether a stepwise approach using microbiota analysis, species-specific quantitative real-time (q)PCRs and culture has the potential to be a more accurate and efficient diagnostic approach than culture alone. Sixty-two sputa obtained in a routine clinical setting from patients with a suspected LRTI were included. All sputa were analysed by culture, microbiota analysis based on the 16S ribosomal RNA gene and multiple species-specific qPCRs. Microbiota and culture data were compared to investigate whether cut-off values for microbiota analysis could be determined. For microbiota analysis, a relative abundance of 25% was identified as the cut-off value for the detection of both genera Streptococcus and Haemophilus. Microbiota analysis combined with species-specific qPCRs resulted in a significant increase in the number of positive sputa (73% vs 58%; p = 0.003) as well as in the number of identified pathogens (51 vs 37; p = 0.049) compared to culture. A stepwise approach using microbiota analysis, species-specific qPCRs and culture has the potential to be used in clinical settings for the diagnosis of LRTIs in the near future.
在临床实践中，下呼吸道感染（LRTIs）的诊断基于培养。本研究旨在评估使用微生物组分析、物种特异性定量实时（q）PCR 和培养的逐步方法是否比单独培养更具准确性和效率。从疑似 LRTI 患者的常规临床环境中获得了 62 份痰液。所有痰液均通过培养、基于 16S 核糖体 RNA 基因的微生物组分析和多种物种特异性 qPCR 进行分析。比较微生物组和培养数据，以确定是否可以确定微生物组分析的截止值。对于微生物组分析，确定 25%的相对丰度为检测链球菌属和嗜血杆菌属的检测截止值。与培养相比，微生物组分析结合物种特异性 qPCR 可显著增加阳性痰液的数量（73%比 58%；p=0.003），以及鉴定出的病原体数量（51 比 37；p=0.049）。使用微生物组分析、物种特异性 qPCR 和培养的逐步方法有可能在不久的将来在临床环境中用于诊断 LRTIs。
