Torres Ignacio, Gimenez Estela, Pascual Tania, Bueno Felipe, Huntley Dixie, Martínez Mireia, Navarro David
Microbiology Service, Hospital Clínico Universitario, Institute for Research INCLIVA, Valencia, Spain.
J Med Microbiol. 2017 Dec;66(12):1752-1758. doi: 10.1099/jmm.0.000643. Epub 2017 Nov 14.
Fast identification of bacteria directly from positive blood cultures (BCs) by matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-TOF MS) can be achieved either using the MALDI Sepsityper kit (protein extraction method) or after a short-term pre-cultivation step on solid medium. We developed a new method that involves short-term enrichment of positive BCs in brain-heart infusion broth (BHI) prior to MALDI-TOF MS analysis.
Eighty-four BCs flagged as positive were included in this study; these were processed in parallel either directly using the MALDI Sepsityper kit or following a short-term culture either in BHI or on Columbia blood agar with 5 % sheep blood (CBA).
Bacterial species were successfully identified in 91.6, 89.2 and 65.4 % of cases after pre-cultivation for 4 h in BHI, on CBA, or by using the MALDI Sepsityper kit, respectively. Overall, the mean incubation time to correct identification was shorter when pre-cultures were performed in BHI; the mean time for Gram-negative rods was 78.2 min in BHI and 108.2 min on CBA (P=0.045), and the mean time for Gram-positive cocci was 128.5 min in BHI and 169.6 min on CBA (P=0.013).
Short-term enrichment of BCs in BHI accelerates identification of a number of bacterial species by MALDI-TOF MS. Further prospective studies are needed to validate our method and gauge its potential clinical impact on the management of bloodstream bacterial infections.
通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)直接从阳性血培养物(BCs)中快速鉴定细菌,可使用MALDI Sepsityper试剂盒(蛋白质提取法),或在固体培养基上进行短期预培养后实现。我们开发了一种新方法,即在MALDI-TOF MS分析之前,先在脑心浸液肉汤(BHI)中对阳性BCs进行短期富集。
本研究纳入了84份标记为阳性的BCs;这些样本被并行处理,要么直接使用MALDI Sepsityper试剂盒,要么在BHI中或含5%绵羊血的哥伦比亚血琼脂(CBA)上进行短期培养。
在BHI中预培养4小时、在CBA上预培养或使用MALDI Sepsityper试剂盒后,分别有91.6%、89.2%和65.4%的病例成功鉴定出细菌种类。总体而言,当在BHI中进行预培养时,正确鉴定的平均孵育时间较短;革兰氏阴性杆菌在BHI中的平均时间为78.2分钟,在CBA上为108.2分钟(P = 0.045),革兰氏阳性球菌在BHI中的平均时间为128.5分钟,在CBA上为169.6分钟(P = 0.013)。
在BHI中对BCs进行短期富集可加速通过MALDI-TOF MS鉴定多种细菌种类。需要进一步的前瞻性研究来验证我们的方法,并评估其对血流细菌感染管理的潜在临床影响。