A multipronged approach for systematic in vitro quantification of catheter-associated biofilms
A multipronged approach for systematic in vitro quantification of catheter-associated biofilms
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DOI：
10.1016/j.hazl.2021.100032
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发表时间：
2021-11
期刊：
影响因子：
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通讯作者：
Alexandra L. Polasko;P. Ramos;R. Kaner;Shaily Mahendra
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文献类型：
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作者：
Alexandra L. Polasko;P. Ramos;R. Kaner;Shaily Mahendra
生物膜是感染的主要原因，特别是那些在医疗管道上发起的感染。病原生物膜的定量对系统确定感染、抗生素处方和植入物置换至关重要。在血液或尿液等液体中检测到传染性病原体时，可能已经形成了大量可能危及生命的生物膜。这些生物膜可以有广泛的病人护理和成本影响。在这里，我们提出了一种非菌株特异性的方案，结合四种微生物测定，这在临床生物膜评估中很少一起使用，以准确量化附着在医疗表面的细胞和细胞外成分。我们的研究结果表明，通过对总蛋白（改良的Lowry）、总生物量（定量聚合酶链反应）、细胞活性（ATP发光）和细胞外聚合物（周期性酸-希夫测定）的联合分析，可以克服标准方法的缺点。采用四种致病的临床分离株（铜绿假单胞菌、金黄色葡萄球菌、耐甲氧西林金黄色葡萄球菌和白色念珠菌）和两种硅胶体外导管验证了这种多管齐下的方法的有效性。尽管感染因子之间的生物膜基质存在差异，但该方法全面量化了从患者获得的留置导管的致病负荷和污垢阻力。该方法具有敏感性、可重复性、多物种特异性和高通量潜力，可用于医院内导管、植入物和呼吸机的质量评估。
Biofilms are a leading cause of infections, especially those initiated on medical tubing. Quantification of pathogenic biofilms is critical for systematic determination of infections, antibiotics prescription, and implant replacement. By the time infectious pathogens are detected in fluids such as blood or urine, substantial and potentially life-threatening biofilms are likely to have already formed. These biofilms can have broad ranging patient care and cost implications. Here we propose a non-strain-specific protocol combining four microbiological assays, which are rarely used together in clinical biofilm assessment, to accurately quantify cellular and extracellular components attached to medical surfaces. Our results demonstrate that the shortcomings of standard approaches can be overcome through the conjunct analysis of total protein (modified Lowry), total biomass (quantitative polymerase chain reaction), cellular activity (ATP luminescence), and extracellular polymeric substances (Periodic Acid-Schiff assay). The efficacy of this multipronged approach was verified using four pathogenic, clinical isolates (Pseudomonas aeruginosa,Staphylococcus aureus, methicillin resistant-Staphylococcus aureus, andCandida albicans) and two types of silicone cathetersin vitro. Despite the variation of biofilm matrices among infectious agents, this approach comprehensively quantified the pathogenic load as well as fouling resistance in indwelling catheters obtained from patients. The sensitivity, reproducibility, multi-species specificity, and high-throughput potential makes this approach valuable for quality assessment of catheters, implants, and ventilators in hospital settings.