Park Jong-Min, Kim Jo-Il, Noh Joo-Yoon, Kim Mira, Kang Min-Jung, Pyun Jae-Chul
Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul, 03722, Republic of Korea.
Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea.
Enzyme Microb Technol. 2017 Feb;97:90-96. doi: 10.1016/j.enzmictec.2016.11.008. Epub 2016 Nov 20.
Many kinds of susceptibility test for β-lactam antibiotics have been used to determine the antibiotic resistance of bacterial strains. Here, a sensitive antibiotic susceptibility test was presented by using a specialized reaction tool for laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS) based on parylene-matrix chip. The β-lactamase assay was carried out in a specialized reaction tool by (1) concentrating the bacterial strain and (2) incubating the bacteria with penicillin-G. The parylene-matrix chip was produced by deposition of a partially porous parylene-N thin film on a dried organic matrix array, and the products of β-lactamase reaction in the low range of mass-to-charge ratio (m/z<500) could be effectively analyzed by using a parylene-matrix chip. The sensing parameters were compared with conventional chromogenic antibiotic susceptibility test for β-lactam antibiotics. Finally, LDI-TOF MS with a specialized reaction tool and parylene-matrix chip could achieve a limit of detection as low as 600 cells/spot for penicillin-G.
许多种β-内酰胺类抗生素敏感性试验已被用于确定细菌菌株的抗生素耐药性。在此，通过使用基于聚对二甲苯基质芯片的激光解吸/电离飞行时间质谱（LDI-TOF MS）专用反应工具，提出了一种灵敏的抗生素敏感性试验。β-内酰胺酶测定在专用反应工具中进行，步骤如下：（1）浓缩细菌菌株；（2）使细菌与青霉素G孵育。聚对二甲苯基质芯片是通过在干燥的有机基质阵列上沉积部分多孔的聚对二甲苯-N薄膜制成的，使用聚对二甲苯基质芯片可以有效分析质荷比范围较低（m/z<500）的β-内酰胺酶反应产物。将传感参数与β-内酰胺类抗生素的传统显色抗生素敏感性试验进行了比较。最后，配备专用反应工具和聚对二甲苯基质芯片的LDI-TOF MS对青霉素G的检测限可低至600个细胞/斑点。
