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焦磷酸测序与基质辅助激光解吸电离飞行时间质谱在慢性肺病细菌鉴定中的性能比较

Performance of pyrosequencing versus MALDI-TOF MS in bacteria identification in chronic lung disease.

作者信息

Navrátilová Lucie, Procházková Petra, Bardoň Jan, Novotný Radko, Zápalka Martin, Jakubec Petr, Zatloukal Jaromír, Kolek Vítězslav, Kopřiva František, Flodrová Pavla, Raclavský Vladislav

机构信息

Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hnevotinska 3, 779 00 Olomouc, Czech Republic.

Laboratory of Growth Regulators, Palacký University Olomouc & Institute of Experimental Botany AS CR, Slechtitelu 27, 783 71 Olomouc, Czech Republic.

出版信息

J Biol Methods. 2016 Aug 13;3(4):e52. doi: 10.14440/jbm.2016.125. eCollection 2016.

Abstract

Rapid identification of the etiological agent in bacterial infection is necessary for correct diagnosis and appropriate therapy. In general, identification of pure cultures of bacteria using conventional phenotyping techniques requires 4-24 hours. Recently available new molecular technologies offer the potential of same day species identification once pure culture is available. Our aim was to evaluate the performance of rDNA V1 hypervariable region pyrosequencing, and the whole cell MALDI-TOF MS protein profiling in routine species identification. During the period from June 2012 to June 2014, 1.140 pure culture isolates were recovered from 402 samples from 126 patients suffering cystic fibrosis, chronic obstructive pulmonary disease or bronchiectasis. All the isolates were subjected to species identification by both techniques. Unfortunately, pyrosequencing was able to reach the species level in 43.2% of isolates only, whereas MALDI-TOF was clearly superior with 96.8% respectively. The overall sensitivity values also clearly underlined the superiority of MALDI-TOF MS with 96.8% compared to 85.1% achieved by pyrosequencing. Generally, MALDI-TOF MS turned out to be the best suitable technique in routine bacterial identification, whereas pyrosequencing could be recommended as the method of choice particularly in situations where MALDI-TOF MS fails to identify rare species.

摘要

快速鉴定细菌感染的病原体对于正确诊断和恰当治疗至关重要。一般来说,使用传统表型分析技术鉴定纯培养细菌需要4到24小时。最近可用的新分子技术提供了在获得纯培养物后当日进行菌种鉴定的可能性。我们的目的是评估rDNA V1高变区焦磷酸测序和全细胞基质辅助激光解吸电离飞行时间质谱蛋白质谱分析在常规菌种鉴定中的性能。在2012年6月至2014年6月期间,从126例患有囊性纤维化、慢性阻塞性肺疾病或支气管扩张症的患者的402份样本中分离出1140株纯培养菌株。所有分离株均采用这两种技术进行菌种鉴定。遗憾的是,焦磷酸测序仅能在43.2%的分离株中鉴定到菌种水平,而基质辅助激光解吸电离飞行时间质谱的鉴定率则高达96.8%,明显更具优势。总体敏感性值也清楚地表明了基质辅助激光解吸电离飞行时间质谱的优越性,其敏感性为96.8%,而焦磷酸测序的敏感性为85.1%。一般来说,基质辅助激光解吸电离飞行时间质谱是常规细菌鉴定中最合适的技术,而焦磷酸测序尤其在基质辅助激光解吸电离飞行时间质谱无法鉴定罕见菌种的情况下可作为首选方法推荐使用。

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