Cueva Angel R, Pham Oanh, Diaby Aissata, Fleming Derek, Rumbaugh Kendra P, Fernandes Gregory E
Department of Surgery and the Burn Center for Research Excellence, Texas Tech University Health Sciences Center, Lubbock, Texas 79430, United States.
Department of Chemical Engineering, Texas Tech University, Lubbock, Texas 79409-3121 United States.
ACS Appl Bio Mater. 2020 Aug 17;3(8):5350-5356. doi: 10.1021/acsabm.0c00665. Epub 2020 Jul 24.
Infection is responsible for up to 60% of deaths in burn patients, and (PA) has the highest mortality rate among all causes of bacteremia. These aggressive and virulent infections are often not detected until there is a positive blood culture for PA, meaning the patient already has bacteremia. An assay that rapidly detects PA while it is still present at low numbers within the burn wound could therefore be transformative, as it would allow for early and more effective treatment intervention. Pyoverdine, an Fe chelating fluorescent molecule, is produced by PA to overcome the problem of limited iron availability and is essential for PA to cause acute infections. Here, we report the development and use of a simple qualitative pyoverdine assay for detection of PA in a clinically relevant burn wound model. In contrast with the 24 h quantitative culture approaches currently used, the pyoverdine assay is complete within 15 min. The assay has a PA detection limit of ∼10 colony forming units/(g of tissue) and detects the PA-specific biomarker pyoverdine within burn wounds before the pathogen disseminates within the body, evidence that the assay can detect lower, prebacteremic levels of PA within burn wounds.
感染是烧伤患者高达60%死亡的原因,而铜绿假单胞菌(PA)在所有菌血症病因中死亡率最高。这些侵袭性和毒性感染往往在PA血培养呈阳性时才被检测到,这意味着患者已经发生菌血症。因此,一种能在PA仍以低数量存在于烧伤创面时快速检测到它的检测方法可能具有变革性,因为它能实现早期且更有效的治疗干预。绿脓菌素是一种铁螯合荧光分子,由PA产生以克服铁供应有限的问题,对PA引起急性感染至关重要。在此,我们报告了一种简单的绿脓菌素定性检测方法的开发及在临床相关烧伤创面模型中检测PA的应用。与目前使用的24小时定量培养方法不同,绿脓菌素检测在15分钟内即可完成。该检测方法对PA的检测限约为10菌落形成单位/(克组织),且在病原体在体内播散之前就能检测到烧伤创面内PA特异性生物标志物绿脓菌素,这证明该检测方法能检测到烧伤创面内PA较低的菌血症前水平。