scholarly journals Color-Coded Droplets and Microscopic Image Analysis for Multiplexed Antibiotic Susceptibility Testing

Biosensors ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 283
Author(s):  
Yunjin Jeong ◽  
Haewook Jang ◽  
Junwon Kang ◽  
Juhong Nam ◽  
Kyoungseob Shin ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Antibiotic Susceptibility ◽  
Drug Testing ◽  
Susceptibility Testing ◽  
Clinical Settings ◽  
Antibiotic Susceptibility Testing ◽  
Testing Conditions ◽  
Clinical Process ◽  
E Coli ◽  
Global Issue

Since the discovery of antibiotics, the emergence of antibiotic resistance has become a global issue that is threatening society. In the era of antibiotic resistance, finding the proper antibiotics through antibiotic susceptibility testing (AST) is crucial in clinical settings. However, the current clinical process of AST based on the broth microdilution test has limitations on scalability to expand the number of antibiotics that are tested with various concentrations. Here, we used color-coded droplets to expand the multiplexing of AST regarding the kind and concentration of antibiotics. Color type and density differentiate the kind of antibiotics and concentration, respectively. Microscopic images of a large view field contain numbers of droplets with different testing conditions. Image processing analysis detects each droplet, decodes color codes, and measures the bacterial growth in the droplet. Testing E. coli ATCC 25922 with ampicillin, gentamicin, and tetracycline shows that the system can provide a robust and scalable platform for multiplexed AST. Furthermore, the system can be applied to various drug testing systems, which require several different testing conditions.

scholarly journals Antibiotic Susceptibility Testing, Plasmid Detection and Curing of Clinically Isolated Enterococcus Species

2019 ◽  
pp. 1-20 ◽  
Author(s):  
G. A. C. Ezeah ◽  
M. C. Ugwu ◽  
A. O. Ekundayo ◽  
O. F. Odo ◽  
O. C. Ike ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Plasmid Dna ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Resistance Pattern ◽  
Antibiotic Susceptibility Testing ◽  
Enterococcus Species ◽  
Transfer Resistance ◽  
Antibiotic Resistance Profile ◽  
Agarose Electrophoresis

Vancomycin resistant enterococci (VRE) are a major medical concern globally. Their significantly greater prevalence and the ability to transfer resistance to vancomycin from other bacteria made them an object of interest and intense research. The isolates of Enterococcus sp. were subjected to antibiotic susceptibility testing before curing. The three Enterococcus species exhibited different antibiotic resistance profile. Pre-curing antibiotic resistance of nosocomial isolates compared with community acquired isolates revealed that high percentage of the nosocomial isolates were resistant to antibiotics compared to community isolate. Post-curing antibiograms of the isolates showed different resistant and susceptibility pattern. Also, DNA plasmid pre-curing and post curing analysis of the isolates showed different resistance pattern. Six of the 15 representative isolates selected on the basis of their high pre-curing antibiotic resistance for plasmid analysis with 0.8% agarose electrophoresis were positive for plasmid DNA. Four (4) of the positive isolates (E. faecium, E. faecium, E. faecalis, and E. avium) had plasmid fragment of greater than 1000 bp while two (2) of them (E. faecalis and E. faecalis) had fragments of between 100 and 500 bp. The remaining nine (9) had no plasmid DNA. The study revealed the pathogenicity factors demonstrated with the enterococcal isolates.

scholarly journals Current and Emerging Methods of Antibiotic Susceptibility Testing

Diagnostics ◽  
2019 ◽  
Vol 9 (2) ◽  
pp. 49 ◽  
Author(s):  
Zeeshan A. Khan ◽  
Mohd F. Siddiqui ◽  
Seungkyung Park
Keyword(s):  
Antibiotic Resistance ◽  
Bacterial Infection ◽  
Economic Burden ◽  
Antibiotic Susceptibility ◽  
Historical Perspective ◽  
Susceptibility Testing ◽  
Empirical Therapy ◽  
Pivotal Role ◽  
Disk Diffusion ◽  
Antibiotic Susceptibility Testing

Antibiotic susceptibility testing (AST) specifies effective antibiotic dosage and formulates a profile of empirical therapy for the proper management of an individual patient’s health against deadly infections. Therefore, rapid diagnostic plays a pivotal role in the treatment of bacterial infection. In this article, the authors review the socio-economic burden and emergence of antibiotic resistance. An overview of the phenotypic, genotypic, and emerging techniques for AST has been provided and discussed, highlighting the advantages and limitations of each. The historical perspective on conventional methods that have paved the way for modern AST like disk diffusion, Epsilometer test (Etest), and microdilution, is presented. Several emerging methods, such as microfluidic-based optical and electrochemical AST have been critically evaluated. Finally, the challenges related with AST and its outlook in the future are presented.

scholarly journals Molecular characterization and antibiotic susceptibility ofVibrio choleraenon-O1

1995 ◽  
Vol 114 (1) ◽  
pp. 51-63 ◽  
Author(s):  
A. Dalsgaard ◽  
O. Serichantalergs ◽  
C. Pitarangsi ◽  
P. Echeverria
Keyword(s):  
Antibiotic Resistance ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Low Molecular Weight ◽  
Toxin Gene ◽  
Antibiotic Susceptibility Testing ◽  
Multiple Antibiotic Resistance ◽  
Heat Stable ◽  
Epidemiological Tool ◽  
Gene Status

SUMMARYA collection of 64 clinical and environmentalVibrio choleraenon-O1 strains isolated in Asia and Peru were characterized by molecular methods and antibiotic susceptibility testing. All strains were resistant to at least 1 and 80% were resistant to two or more antibiotics. Several strains showed multiple antibiotic resistance (≥ three antibiotics). Plasmids most often of low molecular weight were found in 21/64 (33%) strains. The presence of plasmids did not correlate with antibiotic resistance or influence ribotype patterns. In colony hybridization studies 63/64 (98%)V. choleraenon-O1 strains were cholera toxin negative, whereas only strains recovered from patients were heat-stable enterotoxin positive. Forty-sevenBglI ribotypes were observed. No correlation was shown between ribotype and toxin gene status. Ribotype similarity was compared by cluster analysis and two main groups of 13 and 34 ribotypes was found. Ribotyping is apparently a useful epidemiological tool in investigations ofV. choleraenon-O1 infections.

Hospital surveillance predicts community pneumococcal antibiotic resistance in Vietnam

2020 ◽  
Vol 75 (10) ◽  
pp. 2902-2906
Author(s):  
V T T Dai ◽  
J Beissbarth ◽  
P V Thanh ◽  
P T Hoan ◽  
H N L Thuy ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Vaccine Trial ◽  
Nationwide Survey ◽  
Surveillance Data ◽  
Antibiotic Prescribing ◽  
Antibiotic Susceptibility Testing ◽  
Population Demographics ◽  
Testing Data

Abstract Background In Vietnam, Streptococcus pneumoniae is a leading cause of disease, including meningitis. Antibiotics are available without physician prescription at community pharmacies and rates of antibiotic non-susceptibility are high. Appropriate treatment and antibiotic stewardship need to be informed by surveillance data. Objectives To report community-based pneumococcal antibiotic susceptibility testing data from children enrolled in a pneumococcal conjugate vaccine trial in Ho Chi Minh City [the Vietnam Pneumococcal Project (ViPP)] and compare these with published hospital-based data from the nationwide Survey of Antibiotic Resistance (SOAR) to determine whether hospital surveillance data provide an informative estimate of circulating pneumococcal resistance. Methods Pneumococcal isolates from 234 nasopharyngeal swabs collected from ViPP participants at 12 months of age underwent antibiotic susceptibility testing using CLSI methods and the data were compared with SOAR data. Results Antibiotic susceptibility testing identified penicillin-non-susceptible pneumococci in 93.6% of pneumococcus-positive ViPP swabs (oral, non-meningitis breakpoints). Non-susceptibility to erythromycin, trimethoprim/sulfamethoxazole, clindamycin and tetracycline also exceeded 79%. MDR, defined as non-susceptibility to three or more classes of antibiotic, was common (94.4% of swabs). Low or no resistance was detected for ceftriaxone (non-meningitis breakpoints), ofloxacin and vancomycin. Antibiotic non-susceptibility rates in ViPP and SOAR were similar for several antibiotics tested. Conclusions A very high proportion of pneumococci carried in the community are MDR. Despite wide disparities in population demographics between ViPP and SOAR, the non-susceptibility rates for several antibiotics were comparable. Thus, with some qualification, hospital antibiotic susceptibility testing data in Vietnam can inform circulating pneumococcal antibiotic non-susceptibility in young children, the group at highest risk of pneumococcal disease, to guide antibiotic prescribing and support surveillance strategies.

ANTIBIOTIC-RESISTANCE OF HELICOBACTER PYLORI: EPIDEMIOLOGY DIAGNOSIS AND TREATMENT

2012 ◽  
pp. 5-15
Author(s):  
Ngoc Quy Hue Dang ◽  
Van Huy Tran
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Antibiotic Resistance ◽  
Peptic Ulcer ◽  
Peptic Ulcer Disease ◽  
Treatment Regimen ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Antibiotic Susceptibility Testing ◽  
Ulcer Disease

Successful eradicative treatment of H.pylori will reduce the recurrence rate of peptic ulcer disease and the risk of gastric cancer. In our country, the indication of antibiotics for both H.pylori-induced and non-H.pylori-induced diseases has been so available that it has increased H.pylori antibiotic resistance which decreased the efficacy of H.pylori treatment. Some national researches have shown that the rate of H.pylori resistance to Clarithromycin is higher than 20%. For H.pylori-infected patients in whom two empirical regimens fail to eradicate H.pylori, it is necessary to perform culture and antibiogram so that doctors will choose which treatment regimen is suitable for them. Among the H.pylori antibiotic susceptibility testing, the Epsilometer test is both less effort and accurate. H.pylori treatment should be not only standardized and updated according to Maastricht Consensus IV-2012 but also strengthened the management of key antibiotics in H.pylori treatment.

scholarly journals Standardization of Operator-Dependent Variables Affecting Precision and Accuracy of the Disk Diffusion Method for Antibiotic Susceptibility Testing

2015 ◽  
Vol 53 (12) ◽  
pp. 3864-3869 ◽  
Author(s):  
Michael Hombach ◽  
Florian P. Maurer ◽  
Tamara Pfiffner ◽  
Erik C. Böttger ◽  
Reinhard Furrer
Keyword(s):  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Antibiotic Susceptibility Testing ◽  
Content Type ◽  
E Coli ◽  
The Mean ◽  
Precision And Accuracy ◽  
Standard Deviations

Parameters like zone reading, inoculum density, and plate streaking influence the precision and accuracy of disk diffusion antibiotic susceptibility testing (AST). While improved reading precision has been demonstrated using automated imaging systems, standardization of the inoculum and of plate streaking have not been systematically investigated yet. This study analyzed whether photometrically controlled inoculum preparation and/or automated inoculation could further improve the standardization of disk diffusion. Suspensions ofEscherichia coliATCC 25922 andStaphylococcus aureusATCC 29213 of 0.5 McFarland standard were prepared by 10 operators using both visual comparison to turbidity standards and a Densichek photometer (bioMérieux), and the resulting CFU counts were determined. Furthermore, eight experienced operators each inoculated 10 Mueller-Hinton agar plates using a single 0.5 McFarland standard bacterial suspension ofE. coliATCC 25922 using regular cotton swabs, dry flocked swabs (Copan, Brescia, Italy), or an automated streaking device (BD-Kiestra, Drachten, Netherlands). The mean CFU counts obtained from 0.5 McFarland standardE. coliATCC 25922 suspensions were significantly different for suspensions prepared by eye and by Densichek (P< 0.001). Preparation by eye resulted in counts that were closer to the CLSI/EUCAST target of 108CFU/ml than those resulting from Densichek preparation. No significant differences in the standard deviations of the CFU counts were observed. The interoperator differences in standard deviations when dry flocked swabs were used decreased significantly compared to the differences when regular cotton swabs were used, whereas the mean of the standard deviations of all operators together was not significantly altered. In contrast, automated streaking significantly reduced both interoperator differences, i.e., the individual standard deviations, compared to the standard deviations for the manual method, and the mean of the standard deviations of all operators together, i.e., total methodological variation.

scholarly journals Evaluation of an ultra-rapid antibiotic susceptibility testing method on positive blood cultures with Escherichia coli

2021 ◽  
Author(s):  
Özden Baltekin ◽  
Alexander T. A. Johnsson ◽  
Alicia Y. W. Wong ◽  
Kajsa Nilsson ◽  
Bêrivan Mert ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapid Method ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Blood Stream Infection ◽  
Agar Plate ◽  
Blood Cultures ◽  
Antibiotic Susceptibility Testing ◽  
Mortality And Morbidity ◽  
E Coli

Blood stream infection (BSI) is related to high mortality and morbidity. Early antimicrobial therapy is crucial in treating patients with BSI. The most common Gram-negative bacteria causing BSI is Escherichia coli. Targeted effective treatment of patients with BSI is only possible if it is based on antibiotic susceptibility testing (AST) data after blood culture positivity. However, there are very few methods available for rapid phenotypic AST and the fastest method takes 4 h. Here we analyzed the performance of a 30 min ultra-rapid method for AST of E. coli directly from positive blood cultures (BC). In total, 51 positive BC with E. coli were studied, and we evaluated the ultra-rapid method directly on positive BC as well as on E. coli colonies cultured on agar plates. The results obtained by the new method were compared with disk diffusion. The method provided accurate AST result in 30 min to Ciprofloxacin and Gentamicin for 92% and 84% of the positive BC samples, respectively. For E. coli isolates retrieved from agar plates, 86% and 96% of the AST results were accurate for Ciprofloxacin and Gentamicin, respectively, after 30 min of assay time. When time to result was modulated in-silico from 30 to 60 minutes for the agar plate samples, accuracy of AST results went up to 92% for Ciprofloxacin and to 100% for Gentamicin. The present study shows that the method is reliable and delivers ultra-rapid AST data in 30 minutes directly from positive BC and as well as from agar plates.

scholarly journals Multidisciplinary Applied Antibiotic Sensitivity Testing Training in the One Health Approach: ANATOLIAN PROJECT

2021 ◽  
Author(s):  
Serap Süzük Yıldız ◽  
Bunu Kaskatepe ◽  
Özlem Ünaldı ◽  
Hüsniye Şimşek ◽  
Zeynep Ceren Karahan ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Training Program ◽  
Antibiotic Susceptibility ◽  
One Health ◽  
Susceptibility Testing ◽  
Antibiotic Susceptibility Testing ◽  
Sensitivity Testing ◽  
Human Animal ◽  
The One ◽  
Resistance Data

Objective: Antibiotic resistance is one of the most significant problems of human-animal and environmental ecosystems. It is crucial to establish integrated surveillance systems and monitor resistance for the management of antibiotic resistance. Standardization of antibiotic resistance data obtained from various disciplines is the critical point in enhancing the data quality. To realize this objective, a common antibiotic susceptibility testing training program was prepared and performed for professionals from various disciplines to standardize the resistance data to be obtained from the human, animal, and environmental sectors in our country. Method: A total of 48 individuals participated in a five-day training program in three terms. In each period, four small groups, each consisting of four people from a group of different professions, were generated. Participants were trained on quality control, phenotypic tests, genotypic tests, and the use of resistance data in antibiotic susceptibility testing. Pre-test and post-tests were applied to the participants. Results: Individuals with a postgraduate degree who studied antibiotic susceptibility testing in the fields of medicine, veterinary medicine, pharmacy, food, and environment participated in the training. The average number of correct answers in the pre-test and post-test increased from 4.8 to 10.5 in April, from 4 to 9 in June and from 3.4 to 8.5 in September. They studied phenotypic and genotypic tests in the supplied isolates under the supervision of the educators. Conclusion: We presume that dissemination of the training at graduate and postgraduate levels will also enable the One-Health approach to become widespread. In addition, worldwide application of similar trainings will help standardization of resistance data, as well as one health approach.

scholarly journals Whole Slide Imaging for High-Throughput Sensing Antibiotic Resistance at Single-Bacterium Level and Its Application to Rapid Antibiotic Susceptibility Testing

Molecules ◽  
2019 ◽  
Vol 24 (13) ◽  
pp. 2441 ◽  
Author(s):  
Donghui Song ◽  
Haomin Liu ◽  
Huayi Ji ◽  
Yu Lei
Keyword(s):  
Antibiotic Resistance ◽  
High Throughput ◽  
Antibiotic Susceptibility ◽  
Susceptibility Testing ◽  
Large Population ◽  
Accurate Determination ◽  
Great Promise ◽  
Antibiotic Susceptibility Testing ◽  
Whole Slide Imaging ◽  
Antibiotic Resistant

Since conventional culture-based antibiotic susceptibility testing (AST) methods are too time-consuming (typically 24–72 h), rapid AST is urgently needed for preventing the increasing emergence and spread of antibiotic resistant infections. Although several phenotypic antibiotic resistance sensing modalities are able to reduce the AST time to a few hours or less, concerning the biological heterogeneity, their accuracy or limit of detection are limited by low throughput. Here, we present a rapid AST method based on whole slide imaging (WSI)-enabled high-throughput sensing antibiotic resistance at single-bacterium level. The time for determining the minimum inhibitory concentration (MIC) was theoretically shortest, which ensures that the growth of each individual cell present in a large population is inhibited. As a demonstration, our technique was able to sense the growth of at least several thousand bacteria at single-cell level. Reliable MIC of Enterobacter cloacae against gentamicin was obtained within 1 h, while the gold standard broth dilution method required at least 16 h for the same result. In addition, the application of our method prevails over other imaging-based AST approaches in allowing rapid and accurate determination of antibiotic susceptibility for phenotypically heterogeneous samples, in which the number of antibiotic resistant cells was negligible compared to that of the susceptible cells. Hence, our method shows great promise for both rapid AST determination and point-of-care testing of complex clinical bacteria isolates.

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