Department of Chemical Engineering, Northeastern University, Boston, MA, USA.
Department of Civil & Environmental Engineering, Northeastern University, Boston, MA, USA.
Biosens Bioelectron. 2017 Nov 15;97:65-69. doi: 10.1016/j.bios.2017.05.042. Epub 2017 May 24.
In clinical practice, delays in obtaining culture results impact patient care and the ability to tailor antibiotic therapy. Despite the advancement of rapid molecular diagnostics, the use of plate cultures inoculated from swab samples continues to be the standard practice in clinical care. Because the inoculation culture process can take between 24 and 48h before a positive identification test can be run, there is an unmet need to develop rapid throughput methods for bacterial identification. Previous work has shown that pyocyanin can be used as a rapid, redox-active biomarker for identifying Pseudomonas aeruginosa in clinical infections. However, further validation is needed to confirm pyocyanin production occurs in all clinical strains of P. aeruginosa. Here, we validate this electrochemical detection strategy using clinical isolates obtained from patients with hospital-acquired infections or with cystic fibrosis. Square-wave voltammetric scans of 94 different clinical P. aeruginosa isolates were taken to measure the concentration of pyocyanin. The results showed that all isolates produced measureable concentrations of pyocyanin with production rates correlated with patient symptoms and comorbidity. Further bioinformatics analysis confirmed that 1649 genetically sequenced strains (99.9%) of P. aeruginosa possess the two genes (PhzM and PhzS) necessary to produce pyocyanin, supporting the specificity of this biomarker. Confirming the production of pyocyanin by all clinically-relevant strains of P. aeruginosa is a significant step towards validating this strategy for rapid, point-of-care diagnostics.
在临床实践中,获取培养物结果的延迟会影响患者的治疗和调整抗生素治疗的能力。尽管快速分子诊断技术有所进步,但从拭子样本接种的平板培养物仍然是临床护理的标准实践。由于接种培养物的过程需要 24 至 48 小时才能进行阳性鉴定测试,因此需要开发快速高通量的细菌鉴定方法。先前的工作表明,绿脓菌素可以用作鉴定临床感染中铜绿假单胞菌的快速、氧化还原活性生物标志物。然而,需要进一步验证以确认绿脓菌素的产生是否发生在所有临床铜绿假单胞菌菌株中。在这里,我们使用从医院获得性感染或囊性纤维化患者中获得的临床分离株验证了这种电化学检测策略。对 94 种不同的临床铜绿假单胞菌分离株进行方波伏安扫描以测量绿脓菌素的浓度。结果表明,所有分离株均产生可测量浓度的绿脓菌素,其产生率与患者症状和合并症相关。进一步的生物信息学分析证实,1649 种(99.9%)已测序的铜绿假单胞菌菌株具有产生绿脓菌素所需的两个基因(PhzM 和 PhzS),支持了该生物标志物的特异性。确认所有临床相关铜绿假单胞菌菌株均产生绿脓菌素,这是验证该快速即时诊断策略的重要一步。