Abstract:
Phenotypic antimicrobial susceptibility testing enables reliable antibiotic screening but requires multiple strategies to identify each phenotypic change induced by different bactericidal mechanisms. Bacteria apoptosis with typical phenotypic features has never been explored for antibiotic screening. Herein, we developed an antibiotic screening method based on the measurement of antibiotic-induced phosphatidylserine (PS) exposure of apoptotic bacteria. Phosphatidylserine externalization of E. coli that can be widely used as an apoptosis marker for antibiotics with different antibacterial mechanisms was explored. A positively charged PS-binding peptide was immobilized on magnetic beads (MBs) to recognize and capture apoptotic E. coli with PS externalization. Apoptotic E. coli binding led to the charge or charge density change of MBs-peptide, resulting in a potential change on a magneto-controlled polymeric membrane potentiometric sensor. Based on the detection of apoptotic E. coli killed by antibiotics, antibiotic screening for different classes of antibiotics and silver nanoparticles was achieved within 1.5 h using a potentiometric sensor array. This approach enables sensitive, general, and time-saving antibiotic screening, and may open up a new path for antibiotic susceptibility testing.
摘要:
表型抗微生物药敏试验可以进行可靠的抗生素筛选,但需要多种策略来鉴定由不同杀菌机制诱导的每种表型变化。具有典型表型特征的细菌凋亡从未被用于抗生素筛选。在这里,我们基于抗生素诱导的凋亡细菌磷脂酰丝氨酸(PS)暴露量的测定,开发了一种抗生素筛选方法.研究了可广泛用作具有不同抗菌机制的抗生素的凋亡标志物的大肠杆菌磷脂酰丝氨酸外化。将带正电荷的PS结合肽固定在磁珠(MB)上以识别和捕获具有PS外化的凋亡大肠杆菌。凋亡大肠杆菌结合导致MBs-肽的电荷或电荷密度变化,导致磁控聚合物膜电位传感器上的电位变化。基于对抗生素杀死的凋亡大肠杆菌的检测,使用电位传感器阵列在1.5小时内完成了对不同类别抗生素和银纳米颗粒的抗生素筛选。这种方法使敏感,一般,和节省时间的抗生素筛查,并可能为抗生素药敏试验开辟一条新途径。