Department of Chemistry, National Dong Hwa University, Hualien, 974, Taiwan, Republic of China.
Department of Laboratory Medicine and Biotechnology, Tzu Chi University, Hualien, 970, Taiwan, Republic of China.
ChemistryOpen. 2020 Oct 23;9(10):1074-1081. doi: 10.1002/open.202000221. eCollection 2020 Oct.
We present a sensitive and rapid screening method for the determination of β-lactamase activity of antibiotic-resistant bacteria, by designing a pH-sensitive fluorescent dye-doped mesoporous silica nanoparticle encapsulated with penicillin G as a substrate. When penicillin G was hydrolysed by β-lactamase and converted into penicilloic acid, the acidic environment resulted in fluorescence quenching of the dye. The dye-doped mesoporous nanoparticles not only enhanced the β-lactamase-catalyzed reaction rate but also stablized the substrate, penicillin G, which degrades into penicilloic acid in a water solution without β-lactamase. Twentyfive clinical bacterial samples were tested and the antibiotic resistant and susceptible strains were identified. The proposed method may detect the presence of β -lactamases of clinically relevant samples in less than 1 hour. Moreover, the detection limit of β-lactamase activity was as low as 7.8×10 U/mL, which was determined within two hours.
我们提出了一种灵敏快速的筛选方法,用于测定抗生素耐药细菌的β-内酰胺酶活性,方法是设计一种 pH 敏感的荧光染料掺杂介孔硅纳米颗粒,其中包裹有青霉素 G 作为底物。当β-内酰胺酶将青霉素 G 水解转化为青霉素酸时,酸性环境会导致染料荧光猝灭。这种荧光染料掺杂的介孔纳米颗粒不仅增强了β-内酰胺酶催化反应的速率,还稳定了底物青霉素 G,使其在没有β-内酰胺酶的水溶液中降解为青霉素酸。我们对 25 个临床细菌样本进行了测试,并鉴定了抗生素耐药和敏感菌株。该方法可在不到 1 小时的时间内检测到临床相关样本中β-内酰胺酶的存在。此外,β-内酰胺酶活性的检测限低至 7.8×10 U/mL,在两小时内即可确定。