Droplet‐Based Single‐Cell Measurements of 16S rRNA Enable Integrated Bacteria Identification and Pheno‐Molecular Antimicrobial Susceptibility Testing from Clinical Samples in 30 min

BACKGROUND Non fermenting gram-negative bacilli (NFGNB) are a group of heterogenous, aerobic and non-sporing saprophytic bacteria, found as commensals in humans and other animals primarily causing opportunistic healthcare-associated infections. They are innately resistant to many antibiotics and are known to acquire resistance by various mechanisms. They pose a particular difficulty for the healthcare community because multidrug resistance is common and increasing among them and a number of strains have now been identified that exhibit pan drug resistance. This study was conducted to isolate and identify various non-fermenter gram negative bacilli (NFGNB), to study their antibiotic sensitivity pattern and their clinical significance from various clinical samples. METHODS A study was undertaken from March 2019 to February 2020 to isolate NFGNB from various clinical samples received for culture and sensitivity in the department of microbiology in a tertiary care hospital, Ahmedabad. Non lactose fermenting colonies on MacConkey agar plates were further processed by Vitek 2 to identify them and to study their antimicrobial susceptibility testing (AST). RESULTS A total of 2010 NFGNB were isolated from various clinical samples and their AST was evaluated by Vitek 2. Pseudomonas aeruginosa (52.7 %) and Acinetobacter baumannii (36.5 %) were the most common NFGNB isolated. Carbapenem resistance was 93 % for Acinetobacter species and 61 % for Pseudomonas species. CONCLUSIONS Accurate and rapid identification and antimicrobial susceptibility testing of NFGNB help in early initiation of appropriate antimicrobial therapy and proper management of patients thereby help in reducing emergence of MDR strains of NFGNB, mortality and overall hospital stay. KEYWORDS NFGNB – Non-Fermenting Gram-Negative Bacilli, Multidrug Resistance, Pan Drug Resistance, Carbapenem Resistance

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Direct Antimicrobial Susceptibility Testing on Clinical Urine Samples by Optical Tracking of Single Cell Division Events

Small ◽

10.1002/smll.202004148 ◽

2020 ◽

Vol 16 (52) ◽

pp. 2004148

Author(s):

Fenni Zhang ◽

Jiapei Jiang ◽

Michelle McBride ◽

Yunze Yang ◽

Manni Mo ◽

...

Keyword(s):

Cell Division ◽

Single Cell ◽

Antimicrobial Susceptibility ◽

Susceptibility Testing ◽

Antimicrobial Susceptibility Testing ◽

Optical Tracking ◽

Urine Samples

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A Biosensor Platform for Rapid Antimicrobial Susceptibility Testing Directly From Clinical Samples

The Journal of Urology ◽

10.1016/j.juro.2010.09.022 ◽

2011 ◽

Vol 185 (1) ◽

pp. 148-153 ◽

Cited By ~ 60

Author(s):

Kathleen E. Mach ◽

Ruchika Mohan ◽

Ellen Jo Baron ◽

Mei-Chiung Shih ◽

Vincent Gau ◽

...

Keyword(s):

Antimicrobial Susceptibility ◽

Susceptibility Testing ◽

Antimicrobial Susceptibility Testing ◽

Clinical Samples

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Antimicrobial susceptibility testing and identification of exoS and exoU toxin genes of Pseudomonas aeruginosa isolated from clinical samples in lebanon. (c2008)

10.26756/th.2008.46 ◽

2008 ◽

Author(s):

Nahla G. Issa

Keyword(s):

Pseudomonas Aeruginosa ◽

Antimicrobial Susceptibility ◽

Susceptibility Testing ◽

Antimicrobial Susceptibility Testing ◽

Clinical Samples ◽

Toxin Genes

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Nanotube assisted microwave electroporation for single cell pathogen identification and antimicrobial susceptibility testing

Nanomedicine Nanotechnology Biology and Medicine ◽

10.1016/j.nano.2019.01.015 ◽

2019 ◽

Vol 17 ◽

pp. 246-253 ◽

Cited By ~ 7

Author(s):

Jian Gao ◽

Hui Li ◽

Peter Torab ◽

Kathleen E. Mach ◽

David W. Craft ◽

...

Keyword(s):

Single Cell ◽

Antimicrobial Susceptibility ◽

Susceptibility Testing ◽

Antimicrobial Susceptibility Testing ◽

Pathogen Identification

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A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections

Biosensors ◽

10.3390/bios11080288 ◽

2021 ◽

Vol 11 (8) ◽

pp. 288

Author(s):

Britney Forsyth ◽

Peter Torab ◽

Jyong-Huei Lee ◽

Tyler Malcom ◽

Tza-Huei Wang ◽

...

Keyword(s):

Single Cell ◽

Antimicrobial Susceptibility ◽

Susceptibility Testing ◽

Rapid Determination ◽

Bloodstream Infections ◽

Antimicrobial Susceptibility Testing ◽

Resistant Bacteria ◽

Bloodborne Pathogens ◽

The Matrix ◽

Single Cell Trapping

Bloodstream infections are a significant cause of morbidity and mortality worldwide. The rapid initiation of effective antibiotic treatment is critical for patients with bloodstream infections. However, the diagnosis of bloodborne pathogens is largely complicated by the matrix effect of blood and the lengthy blood tube culture procedure. Here we report a culture-free workflow for the rapid isolation and enrichment of bacterial pathogens from whole blood for single-cell antimicrobial susceptibility testing (AST). A dextran sedimentation step reduces the concentration of blood cells by 4 orders of magnitude in 20–30 min while maintaining the effective concentration of bacteria in the sample. Red blood cell depletion facilitates the downstream centrifugation-based enrichment step at a sepsis-relevant bacteria concentration. The workflow is compatible with common antibiotic-resistant bacteria and does not influence the minimum inhibitory concentrations. By applying a microfluidic single-cell trapping device, we demonstrate the workflow for the rapid determination of bacterial infection and antimicrobial susceptibility testing at the single-cell level. The entire workflow from blood to categorical AST result can be completed in less than two hours.

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Multiplexed Automated Digital Microscopy for Rapid Identification and Antimicrobial Susceptibility Testing of Bacteria and Yeast Directly from Clinical Samples

Clinical Microbiology Newsletter ◽

10.1016/j.clinmicnews.2015.10.001 ◽

2015 ◽

Vol 37 (20) ◽

pp. 161-167 ◽

Cited By ~ 17

Author(s):

Christina Chantell

Keyword(s):

Antimicrobial Susceptibility ◽

Susceptibility Testing ◽

Rapid Identification ◽

Antimicrobial Susceptibility Testing ◽

Clinical Samples ◽

Digital Microscopy

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Taxonomy, virulence determinants and antimicrobial susceptibility of Aeromonas spp. isolated from bacteremia in southeastern China

Antimicrobial Resistance and Infection Control ◽

10.1186/s13756-021-00911-0 ◽

2021 ◽

Vol 10 (1) ◽

Author(s):

Yao Sun ◽

Yajie Zhao ◽

Wenya Xu ◽

Renchi Fang ◽

Qing Wu ◽

...

Keyword(s):

Antimicrobial Susceptibility ◽

Poor Prognosis ◽

Susceptibility Testing ◽

Clinical Manifestations ◽

Virulence Gene ◽

Antimicrobial Susceptibility Testing ◽

Clinical Samples ◽

Virulence Determinants ◽

Southeastern China ◽

Effective Measure

Abstract Background The study aimed to elucidate the species taxonomy, clinical manifestations, virulence gene profiles and antimicrobial susceptibilities of Aeromonas strains isolated from life-threatening bacteremia in southeastern China. Methods Clinical samples of Aeromonas causing bacteremia were isolated from a teaching hospital in Wenzhou from 2013 to 2018 and a retrospective cohort study was performed. Aeromonas strains were identified at species level by housekeeping gene gyrB. Virulence and drug resistance-associated genes were screened by polymerase chain reaction (PCR) and antimicrobial susceptibility testing (AST) was performed by the VITEK 2 Compact system. Results A total of 58 Aeromonas isolated from patients with bacteremia were collected during 6 years (2013–2018). 58 isolates were identified to five different species, where Aeromonas dhakensis appeared to be the predominant species (26/58), followed by Aeromonas veronii (13/58), Aeromonas caviae (10/58), Aeromonas hydrophila (7/58) and Aeromonas jandaei (2/58). 16 of 58 patients had poor prognosis. Poor prognosis was significantly associated with liver cirrhosis and inappropriate empirical antimicrobials therapy. The progression of bacteremia caused by Aeromonas was extremely fast, especially in A. dhakensis infections. Virulence genes aer, lip, hlyA, alt, ast, and act, were detected at ratios of 24.1% (14/58), 62.1% (36/58), 65.5% (38/58), 58.6% (34/58), 15.5% (9/58) and 65.5% (38/58), respectively. Antimicrobial susceptibility testing exhibited that 9 out of 58 isolates were identified as multi-drug resistant (MDR) organism. The blaTEM gene was identified in all 9 MDR isolates. blaSHV, blaAQU-1, blaMOX, blaCepH, blaCphA and aac(6′)-Ib-cr were detected in 4 isolates, 2 isolates, 1 isolate, 3 isolates, 8 isolates, and 3 isolates, respectively. The majority of Aeromonas strains maintained susceptible to 3rd generation cephalosporins, aminoglycosides, fluoroquinolones and furantoin. Conclusions The prevalence and dangerousness of Aeromonas infections, especially A. dhakensis, are underestimated in clinic. Continuous monitoring is essential to keep track of MDR Aeromonas due to the increasing prevalence recently and a more effective measure is required to control the spread of resistance determinants.

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