Hypersensitive antibiotic susceptibility test based on a β-lactamase assay with a parylene-matrix chip

• Published in: Enzyme and microbial technology Vol. 97; pp. 90 - 96
• Main Authors: Park, Jong-Min, Kim, Jo-Il, Noh, Joo-Yoon, Kim, Mira, Kang, Min-Jung, Pyun, Jae-Chul
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.02.2017
• Subjects: Anti-Bacterial Agents - pharmacology; Antibiotic susceptibility test; beta-Lactamase Inhibitors - pharmacology; beta-Lactamases - analysis; beta-Lactams - pharmacology; Biotechnology; Escherichia coli - drug effects; Escherichia coli - enzymology; Laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS); Limit of Detection; Microbial Sensitivity Tests - instrumentation; Microbial Sensitivity Tests - methods; Microbial Sensitivity Tests - statistics & numerical data; Parylene-matrix chip; Polymers; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Xylenes; β-lactamase; Parylene-matrix chip; Antibiotic susceptibility test; Laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS); β-lactamase
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/j.enzmictec.2016.11.008

[Display omitted] •A reaction tool was developed for β-lactamase assay with bacterial strains.•The reaction tool was used for effectively concentrated reaction products.•Reaction products were analyzed using LDI-TOF MS with parylene-matrix chip.•Finally, the antibiotics susceptibility test with LOD of 600 cells/spot was achieved. 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.