Multi-screening of beta-lactam antibiotics for beta-lactamase resistance by means of a paper-based analytical device with a 4-(2-pyridylazo)resorcinol (PAR)-Hg2+ complex

Abstract

beta-Lactam antibiotics are widely used for the treatment of bacterial infections; however, an increase in the number of antibiotic-resistant bacteria, which produce beta-lactamases that inactivate beta-lactam antibiotics, can lead to serious untreatable diseases. Thus, proper prescribing of antibiotics and the development of new antimicrobial agents with strong resistance to hydrolysis by beta-lactamases can be a solution to reduce antibiotic resistance and treat various bacterial infections. Accordingly, simple and rapid methods for the screening of antibiotics that show beta-lactamase resistance are required. Therefore, we designed a multi-channel microfluidic paper-based analytical device (mPAD) for colorimetric screening of beta-lactam antibiotics for beta-lactamase resistance; the mPAD, created by modifying paper with self-assembling organosilane and a 4-(2-pyridylazo) resorcinol (PAR)-Hg2+ complex, can screen beta-lactam antibiotics for hydrolytic resistance to multiple beta-lactamases at once. To demonstrate its suitability for this task, we confirmed the resistance of already studied beta-lactam antibiotics-penicillin G, cloxacillin, and ceftazidime-toward different classes of beta-lactamases (penicillinase, TEM-1, and CTX-M-1) using the mPAD. Based on ligand exchange between hydrolyzed beta-lactam antibiotics and PAR in the PAR-Hg2+ complex, colorimetric detection of the relative susceptibilities of beta-lactam antibiotics to a beta-lactamase is possible even with the naked eye in only 10 minutes. The portable and equipment-free mPAD allows for simple multi-screening of diverse classes of beta-lactam antibiotics for beta-lactamase resistance.