Very rapid flow cytometric assessment of antimicrobial susceptibility during the apparent lag phase of microbial (re)growth
Very rapid flow cytometric assessment of antimicrobial susceptibility during the apparent lag phase of microbial (re)growth
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DOI：
10.1101/480392
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发表时间：
2018-11
期刊：
影响因子：
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通讯作者：
Srijan Jindal;H. Thampy;P. Day;D. Kell
中科院分区：
文献类型：
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作者：
Srijan Jindal;H. Thampy;P. Day;D. Kell
大肠杆菌细胞在LB培养基中生长，从2-4h的固定相中取出，然后重新接种到浓度为105的新鲜培养基中。mL-1或更低)细菌尿的特征。流式细胞术用于评估我们检测细胞大小、数量、膜激活（使用碳氰染料）和DNA分布的变化的速度。结果表明，虽然通过体积外径测量宏观观察到的滞后期长达4h，但真正的滞后期可能小于15- 20min，并伴有许多可观察到的生化变化。细胞敏感的抗生素在重新接种20分钟内影响了这些变化，提供了一种非常快速的抗生素敏感性测试的可能性，与去全科医生诊所的时间一致。该策略成功应用于从医生手术中获取的真正潜在尿路感染（UTI）样本。所开发的方法可以证明在确保正确的处方从而降低抗菌素耐药性的传播方面具有相当大的价值。
Cells of E. coli were grown in LB medium, taken from a stationary phase of 2-4h, and reinoculated into fresh media at a concentration (105.mL-1 or lower) characteristic of bacteriuria. Flow cytometry was used to assess how quickly we could detect changes in cell size, number, membrane energisation (using a carbocyanine dye) and DNA distribution. It turned out that while the lag phase observable macroscopically via bulk OD measurements could be as long as 4h, the true lag phase could be less than 15-20 min, and was accompanied by many observable biochemical changes. Antibiotics to which the cells were sensitive affected these changes within 20 min of reinoculation, providing the possibility of a very rapid antibiotic susceptibility test, on a timescale compatible with a visit to a GP clinic. The strategy was applied successfully to genuine potential Urinary Tract Infection (UTI) samples taken from a doctor’s surgery. The methods developed could prove of considerable value in ensuring the correct prescription and thereby lowering the spread of antimicrobial resistance.