Gato Eva, Constanso Ignacio Pedro, Rodiño-Janeiro Bruno Kotska, Guijarro-Sánchez Paula, Alioto Tyler, Arroyo Manuel Jesús, Méndez Gema, Mancera Luis, Gut Marta, Gut Ivo, Álvarez-Tejado Miguel, Bou Germán, Oviaño Marina
Servicio de Microbiología, Complejo Hospitalario Universitario A Coruña, A Coruña, Spain.
Servicio de Análisis Clínicos. Complejo Hospitalario Universitario A Coruña, A Coruña, Spain.
J Clin Microbiol. 2021 Jul 19;59(8):e0023821. doi: 10.1128/JCM.00238-21.
Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been used for the direct detection of KPC-producing isolates by analysis of the 11,109 Da mass peak representing the P019 protein. In this study, we evaluate the presence of the 11,109 Da mass peak in a collection of 435 unduplicated Klebsiella pneumoniae clinical isolates. The prevalence of the P019 peak in the K. pneumoniae isolates was 49.2% (32/65). The 11,109 Da mass peak was not observed in any of the other carbapenemase (319) or noncarbapenemase producers (116). Computational analysis of the presence of the gene was performed in the aforementioned carbapenemase-producing K. pneumoniae isolates fully characterized by whole-genome sequencing (WGS) and in a further collection of 1,649 K. pneumoniae genomes included in EuSCAPE. Herein, we have demonstrated that the gene is not exclusively linked to the pKpQil plasmid but that it is present in the following plasmids: IncFIB(K)/IncFII(K)/ColRNAI, IncFIB(pQil), IncFIB(pQil)/ColRNAI, IncFIB(pQil)/IncFII(K), IncFIB(K)/IncFII(K), and IncX3. In addition, we have proven the independent movement of the Tn and the IS, of which the gene is a component. The absence of the gene was obvious in Col440I, Col(pHAD28), IncFIB(K)/IncX3/IncFII(K), and IncFIB(K)/IncFII(K) plasmids. In addition, we also observed another plasmid in which neither Tn nor IS was found, IncP6. In the EuSCAPE, the occurrence of varied from 0% to 100% among the different geographical locations. The adverse clinical impact of the diminished prevalence of the gene within the plasmid encoding KPC-producing Klebsiella pneumoniae puts forward the need for reconsideration when applying this technique in a clinical setting.
基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）最近已用于通过分析代表P019蛋白的11,109 Da质量峰直接检测产KPC的分离株。在本研究中，我们评估了435株未重复的肺炎克雷伯菌临床分离株中11,109 Da质量峰的存在情况。肺炎克雷伯菌分离株中P019峰的流行率为49.2%（32/65）。在任何其他碳青霉烯酶产生菌（319株）或非碳青霉烯酶产生菌（116株）中均未观察到11,109 Da质量峰。对上述通过全基因组测序（WGS）完全鉴定的产碳青霉烯酶肺炎克雷伯菌分离株以及EuSCAPE中另外1649株肺炎克雷伯菌基因组进行了该基因存在情况的计算分析。在此，我们证明该基因并非仅与pKpQil质粒相关，而是存在于以下质粒中：IncFIB(K)/IncFII(K)/ColRNAI、IncFIB(pQil)、IncFIB(pQil)/ColRNAI、IncFIB(pQil)/IncFII(K)、IncFIB(K)/IncFII(K)和IncX3。此外，我们证实了Tn和IS（该基因是其组成部分）的独立移动。在Col440I、Col(pHAD28)、IncFIB(K)/IncX3/IncFII(K)和IncFIB(K)/IncFII(K)质粒中明显不存在该基因。此外，我们还观察到另一种既未发现Tn也未发现IS的质粒IncP6。在EuSCAPE中，不同地理位置的该基因出现率从0%到100%不等。编码产KPC肺炎克雷伯菌的质粒中该基因流行率降低所带来的不良临床影响，提出了在临床环境中应用该技术时需要重新考虑的必要性。
