Idelevich E A, Grünastel B, Peters G, Becker K
Institute of Medical Microbiology, University Hospital Münster, Germany.
New Microbes New Infect. 2015 Dec 23;10:19-24. doi: 10.1016/j.nmni.2015.12.004. eCollection 2016 Mar.
Pathogen identification and antimicrobial susceptibility testing (AST) should be available as soon as possible for patients with bloodstream infections. We investigated whether a lysis-centrifugation (LC) blood culture (BC) method, combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification and Vitek 2 AST, provides a time advantage in comparison with the currently used automated broth-based BC system. Seven bacterial reference strains were added each to 10 mL human blood in final concentrations of 100, 10 and 1 CFU/mL. Inoculated blood was added to the Isolator 10 tube and centrifuged at 3000 g for 30 min, then 1.5 mL sediment was distributed onto five 150-mm agar plates. Growth was observed hourly and microcolonies were subjected to MALDI-TOF MS and Vitek 2 as soon as possible. For comparison, seeded blood was introduced into an aerobic BC bottle and incubated in the BACTEC 9240 automated BC system. For all species/concentration combinations except one, successful identification and Vitek 2 inoculation were achieved even before growth detection by BACTEC. The fastest identification and inoculation for AST were achieved with Escherichia coli in concentrations of 100 CFU/mL and 10 CFU/mL (after 7 h each, while BACTEC flagged respective samples positive after 9.5 h and 10 h). Use of the LC-BC method allows skipping of incubation in automated BC systems and, used in combination with rapid diagnostics from microcolonies, provides a considerable advantage in time to result. This suggests that the usefulness of direct BC on solid medium should be re-evaluated in the era of rapid microbiology.
对于血流感染患者,应尽快进行病原体鉴定和抗菌药物敏感性试验(AST)。我们研究了裂解离心(LC)血培养(BC)方法结合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定和Vitek 2 AST,与目前使用的基于肉汤的自动化BC系统相比是否具有时间优势。将七种细菌参考菌株分别以100、10和1 CFU/mL的终浓度加入10 mL人血中。将接种的血液加入Isolator 10管中,以3000 g离心30分钟,然后将1.5 mL沉淀物分布在五个150-mm琼脂平板上。每小时观察生长情况,一旦有微菌落,尽快对其进行MALDI-TOF MS和Vitek 2检测。为作比较,将接种的血液接种到需氧BC瓶中,并在BACTEC 9240自动化BC系统中培养。除一种情况外对于所有物种/浓度组合,甚至在BACTEC检测到生长之前就成功进行了鉴定和Vitek 2接种。对于浓度为100 CFU/mL和10 CFU/mL的大肠杆菌,AST的鉴定和接种速度最快(每种情况均在7小时后完成,而BACTEC分别在9.5小时和10小时后标记相应样本为阳性)。使用LC-BC方法可跳过在自动化BC系统中的培养,并且与来自微菌落的快速诊断相结合,在获得结果的时间上具有相当大的优势。这表明在快速微生物学时代应重新评估在固体培养基上直接进行血培养的实用性。