Novel Electrical Method for the Rapid Determination of Minimum Inhibitory Concentration (MIC) and Assay of Bactericidal/Bacteriostatic Activity

Puttaswamy, Sachidevi; Lee, Byung-Doo; Amighi, Banoo; Chakraborty, Suman Bhusan; Sengupta, Shramik

doi:10.4172/2155-6210.s2-003

Cited by 11 publications

(9 citation statements)

References 28 publications

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“…Gas-permeable polydimethylsiloxane (PDMS) microchannels with high surface-to-volume ratios have also been demonstrated to perform AST in 2 h. 12 To enable point-of-care applications, an electrochemical biosensor has been demonstrated to determine the pathogen antibiotic resistance profiles from clinical urine samples within 3.5 h. 13 Furthermore, impedance spectroscopy can also be applied to detect the bactericidal and bacteriostatic effect of antibiotics. 14 Recently, single-cell AST approaches with agarose or microfluidic confinements have also been reported. 15,16 An alternative approach to avoid bulky, expensive optical equipment is to develop cell phone–based diagnostic systems.…”

Section: Introductionmentioning

confidence: 99%

A Cell Phone–Based Microphotometric System for Rapid Antimicrobial Susceptibility Testing

et al. 2014

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This study demonstrates a low-cost, portable diagnostic system for rapid antimicrobial susceptibility testing in resource-limited settings. To determine the antimicrobial resistance phenotypically, the growth of pathogens in microwell arrays is detected under different antibiotic conditions. The use of a colorimetric cell viability reagent is shown to significantly improve the sensitivity of the assay compared with standard absorbance spectroscopy. Gas-permeable microwell arrays are incorporated for facilitating rapid bacterial growth and eliminating the requirement of bulky supporting equipment. Antibiotics can also be precoated in the microwell array to simplify the assay protocol toward point-of-care applications. Furthermore, a low-cost cell phone–based microphotometric system is developed for detecting the bacterial growth in the microwell array. By optimizing the operating conditions, the system allows antimicrobial susceptibility testing for samples with initial concentrations from 101 to 106 cfu/mL. Using urinary tract infection as the model system, we demonstrate rapid antimicrobial resistance profiling for uropathogens in both culture media and urine. With its simplicity and cost-effectiveness, the cell phone–based microphotometric system is anticipated to have broad applicability in resource-limited settings toward the management of infectious diseases caused by multidrug-resistant pathogens.

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Section: Introductionmentioning

confidence: 99%

A Cell Phone–Based Microphotometric System for Rapid Antimicrobial Susceptibility Testing

et al. 2014

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“…For our technique, at regular intervals of time, small aliquots of sample are drawn and its impedance is tested using impedance analyzer. The data is fitted to an equivalent circuit to generate a Nyquist plot [ 48 ] and thereby estimates the values of the desired circuit parameters. We show on the graph three sets of data: those obtained at 2, 8, and 12 h for a culture of M. smegmatis ( large , medium , and small semi-circles ).…”

Section: Methodsmentioning

confidence: 99%

“…The basic principles governing the use of m-EIS to detect microorganisms have been described in our prior work [ 36 , 37 , 48 ]. Briefly, the protocol requires us to periodically (every 12–24 h for M. bovis BCG , and 2–4 h for M. smegmatis ) perform an electrical “scan” of sample aliquots in a microfluidic cassette, wherein we measure electrical impedance at multiple (200) frequencies ranging from 1 kHz to 100 MHz.…”

Section: Methodsmentioning

confidence: 99%

Rapid culture-based detection of living mycobacteria using microchannel electrical impedance spectroscopy (m-EIS)

et al. 2017

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BackgroundMultiple techniques exist for detecting Mycobacteria, each having its own advantages and drawbacks. Among them, automated culture-based systems like the BACTEC-MGIT™ are popular because they are inexpensive, reliable and highly accurate. However, they have a relatively long “time-to-detection” (TTD). Hence, a method that retains the reliability and low-cost of the MGIT system, while reducing TTD would be highly desirable.MethodsLiving bacterial cells possess a membrane potential, on account of which they store charge when subjected to an AC-field. This charge storage (bulk capacitance) can be estimated using impedance measurements at multiple frequencies. An increase in the number of living cells during culture is reflected in an increase in bulk capacitance, and this forms the basis of our detection. M. bovis BCG and M. smegmatis suspensions with differing initial loads are cultured in MGIT media supplemented with OADC and Middlebrook 7H9 media respectively, electrical “scans” taken at regular intervals and the bulk capacitance estimated from the scans. Bulk capacitance estimates at later time-points are statistically compared to the suspension’s baseline value. A statistically significant increase is assumed to indicate the presence of proliferating mycobacteria.ResultsOur TTDs were 60 and 36 h for M. bovis BCG and 20 and 9 h for M. smegmatis with initial loads of 1000 CFU/ml and 100,000 CFU/ml respectively. The corresponding TTDs for the commercial BACTEC MGIT 960 system were 131 and 84.6 h for M. bovis BCG and 41.7 and 12 h for M smegmatis, respectively.ConclusionOur culture-based detection method using multi-frequency impedance measurements is capable of detecting mycobacteria faster than current commercial systems.

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“…Thus, the concentration of the antibiotic at which no change in the signal is seen over time or when a decrease in the signal is observed, is considered their MIC. Since the system does not need to wait to see bacterial growth-which is limited by the doubling time of the bacteria, but detect bacterial death in the presence of antibiotics, it can lead to faster AST profiles (<4 hours) [30]. The method can also distinguish between bactericidal and bacteriostatic effect of the antibiotics on samples and can go directly from samples without need for pure isolates.…”

Section: Microfluidic Electrical Impedance Spectroscopy (M-eis) Systemmentioning

confidence: 99%

A Comprehensive Review of the Present and Future Antibiotic Susceptibility Testing (AST) Systems

Puttaswamy¹,

Gupta²,

Regunath³

et al. 2018

Arch Clin Microbiol

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Tools and instruments available in the clinical microbiology labs for analysis of patient samples and diagnosis are constantly evolving. The main impetus behind this is to decrease the overall time taken to obtain the results from the instruments, enhance the ease of sample processing, increasing the sample turn-around time with the ultimate goal of earlier patient treatment and better recovery rates. This is especially true in the case of antibiotic susceptibility testing (AST), where every hour saved in obtaining the results leading to an earlier switch to targeted antibiotic therapy will have a direct influence on improving clinical outcomes. Reduction in the time to obtain AST results reduces the duration of use of broad-spectrum antibiotics, which in turn decreases the emergence of antibiotic resistance among bacteria. Many of the traditional methods available for AST are labor intensive and slow despite being precise in obtaining results. Thus, there is a trend towards development and use of automated diagnostic devices which are rapid and easy to use. This review article provides a detailed summary of traditional AST methods, currently used automated methods, and focuses on some of the promising emerging and future technologies in the field of rapid antibiotic susceptibility profiling.

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Novel Electrical Method for the Rapid Determination of Minimum Inhibitory Concentration (MIC) and Assay of Bactericidal/Bacteriostatic Activity

Cited by 11 publications

References 28 publications

A Cell Phone–Based Microphotometric System for Rapid Antimicrobial Susceptibility Testing

A Cell Phone–Based Microphotometric System for Rapid Antimicrobial Susceptibility Testing

Rapid culture-based detection of living mycobacteria using microchannel electrical impedance spectroscopy (m-EIS)

A Comprehensive Review of the Present and Future Antibiotic Susceptibility Testing (AST) Systems

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