Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Università di Pisa, Via San Zeno 37, 56127, Pisa, Italy.
BMC Microbiol. 2019 Nov 29;19(1):267. doi: 10.1186/s12866-019-1641-1.
The application of matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) to microbial identification has allowed the development of rapid methods for identification of microorganisms directly in positive, blood cultures (BCs). These methods can yield accurate results for monomicrobial BCs, but often fail to identify multiple microorganisms in polymicrobial BCs. The present study was aimed at establishing a rapid and simple method for identification of bacteria and yeast in polymicrobial BCs from patients with bloodstream infection.
The rapid method herein proposed is based on short-term culture in liquid media allowing selective growth of microorganisms recovered from polymicrobial BCs, followed by rapid identification by MALDI-TOF MS. To evaluate the accuracy of this method, 56 polymicrobial BCs were comparatively analyzed with the rapid and routine methods. The results showed concordant identification for both microbial species in 43/50 (86%) BCs containing two different microorganisms, and for two microbial species in six BCs containing more than two different species. Overall, 102/119 (85.7%) microorganisms were concordantly identified by the rapid and routine methods using a cut-off value of 1.700 for valid identification. The mean time to identification after BC positivity was about 4.2 h for streptococci/enterococci, 8.7 h for staphylococci, 11.1 h for Gram-negative bacteria, and 14.4 h for yeast, allowing a significant time saving compared to the routine method.
The proposed method allowed rapid and reliable microbial identification in polymicrobial BCs, and could provide clinicians with timely, useful information to streamline empirical antimicrobial therapy in critically ill patients.
基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱(MS)在微生物鉴定中的应用,使得直接从阳性、血培养物(BC)中鉴定微生物的快速方法得以发展。这些方法对于单微生物 BC 可以得出准确的结果,但往往无法识别多微生物 BC 中的多种微生物。本研究旨在建立一种快速、简单的方法,用于鉴定血流感染患者多微生物 BC 中的细菌和酵母。
本文提出的快速方法基于短期液体培养,允许从多微生物 BC 中回收的微生物选择性生长,然后通过 MALDI-TOF MS 快速鉴定。为了评估该方法的准确性,对 56 份多微生物 BC 进行了快速法和常规法的比较分析。结果显示,在包含两种不同微生物的 43/50(86%)BC 中,两种微生物物种的鉴定结果均一致,在包含两种以上不同物种的 6 个 BC 中,两种微生物物种的鉴定结果也一致。总体而言,使用 1.700 的截止值,快速法和常规法对 102/119(85.7%)微生物的鉴定结果一致。BC 阳性后,链球菌/肠球菌的鉴定时间约为 4.2 小时,葡萄球菌为 8.7 小时,革兰氏阴性菌为 11.1 小时,酵母为 14.4 小时,与常规方法相比,可显著缩短鉴定时间。
该方法可快速、可靠地鉴定多微生物 BC,为临床医生提供及时、有用的信息,简化重症患者的经验性抗菌治疗。