Highly sensitive bacterial susceptibility test against penicillin using parylene-matrix chip.
This work presented a highly sensitive bacterial antibiotic susceptibility test through β-lactamase assay using Parylene-matrix chip. β-lactamases (EC 3.5.2.6) are an important family of enzymes that confer resistance to β-lactam antibiotics by catalyzing the hydrolysis of these antibiotics. Here we present a highly sensitive assay to quantitate β-lactamase-mediated hydrolysis of penicillin into penicilloic acid. Typically, MALDI-TOF mass spectrometry has been used to quantitate low molecular weight analytes and to discriminate them from noise peaks of matrix fragments that occur at low m/z ratios (m/z<500). The β-lactamase assay for the Escherichia coli antibiotic susceptibility test was carried out using Parylene-matrix chip and MALDI-TOF mass spectrometry. The Parylene-matrix chip was successfully used to quantitate penicillin (m/z: [PEN+H](+)=335.1 and [PEN+Na](+)=357.8) and penicilloic acid (m/z: [PA+H](+)=353.1) in a β-lactamase assay with minimal interference of low molecular weight noise peaks. The β-lactamase assay was carried out with an antibiotic-resistant E. coli strain and an antibiotic-susceptible E. coli strain, revealing that the minimum number of E. coli cells required to screen for antibiotic resistance was 1000 cells for the MALDI-TOF mass spectrometry/Parylene-matrix chip assay.
Department of Materials Science and Engineering Yonsei University, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-749, Republic of Korea.
Jong-Min
JM
Park
Department of Materials Science and Engineering Yonsei University, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-749, Republic of Korea.
Jo-Il
JI
Kim
Department of Materials Science and Engineering Yonsei University, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-749, Republic of Korea.
Hyun-Woo
HW
Song
Department of Materials Science and Engineering Yonsei University, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-749, Republic of Korea.
Joo-Yoon
JY
Noh
Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea.
Min-Jung
MJ
Kang
Department of Materials Science and Engineering Yonsei University, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-749, Republic of Korea. Electronic address: jcpyun@yonsei.ac.kr.
Jae-Chul
JC
Pyun
D001681:Biological Assay / Q000295:instrumentation; D024881:Drug Resistance, Bacterial; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology*; D046228:Microarray Analysis / Q000295:instrumentation; D008826:Microbial Sensitivity Tests / Q000295:instrumentation*; D010397:Penicillanic Acid / Q000031:analogs & derivatives / Q000032:analysis / Q000378:metabolism; D010406:Penicillins / Q000493:pharmacokinetics / Q000494:pharmacology*; D011108:Polymers / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014992:Xylenes / Q000737:chemistry*; D001618:beta-Lactamases / Q000032:analysis / Q000378:metabolism*
D010406:Penicillins; D011108:Polymers; D014992:Xylenes; C008782:penicilloic acid; C011055:parylene; D010397:Penicillanic Acid; D001618:beta-Lactamases
MALDI-TOF mass spectrometry; Parylene-matrix chip; Penicillin; β-lactamase