scholarly journals Emerging Resistance, New Antimicrobial Agents  …  but No Tests! The Challenge of Antimicrobial Susceptibility Testing in the Current US Regulatory Landscape

2016 ◽  
Vol 63 (1) ◽  
pp. 83-88 ◽  
Author(s):  
R. M. Humphries ◽  
J. A. Hindler
Keyword(s):  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Regulatory Landscape

scholarly journals The Frequency of Methicillin-Resistant Staphylococcus aureus and Coagulase Gene Polymorphism in Egypt

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Hend M. Abdulghany ◽  
Rasha M. Khairy
Keyword(s):  
Staphylococcus Aureus ◽  
Gene Polymorphism ◽  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Methicillin Resistant ◽  
Microbiological Methods ◽  
Rflp Typing

The current study aimed to use Coagulase gene polymorphism to identify methicillin-resistant Staphylococcus aureus (MRSA) subtypes isolated from nasal carriers in Minia governorate, Egypt, evaluate the efficiency of these methods in discriminating variable strains, and compare these subtypes with antibiotypes. A total of 400 specimens were collected from nasal carriers in Minia governorate, Egypt, between March 2012 and April 2013. Fifty-eight strains (14.5%) were isolated and identified by standard microbiological methods as MRSA. The identified isolates were tested by Coagulase gene RFLP typing. Out of 58 MRSA isolates 15 coa types were classified, and the amplification products showed multiple bands (1, 2, 3, 4, 5, and 8 bands). Coagulase gene PCR-RFLPs exhibited 10 patterns that ranged from 1 to 8 fragments with AluI digestion. Antimicrobial susceptibility testing with a panel of 8 antimicrobial agents showed 6 different antibiotypes. Antibiotype 1 was the most common phenotype with 82.7%. The results have demonstrated that many new variants of the coa gene are present in Minia, Egypt, different from those reported in the previous studies. So surveillance of MRSA should be continued.

scholarly journals Analysis of Whole-Genome Sequences for the Prediction of Penicillin Resistance and β-Lactamase Activity inBacillus anthracis

mSystems ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
A. S. Gargis ◽  
H. P. McLaughlin ◽  
A. B. Conley ◽  
C. Lascols ◽  
P. A. Michel ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Antimicrobial Susceptibility ◽  
Sigma Factor ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Penicillin Resistance ◽  
Whole Genome ◽  
Activity Levels ◽  
Content Type

ABSTRACTPenicillin (PEN) is a low-cost option for anthrax treatment, but naturally occurring resistance has been reported. β-Lactamase expression (bla1,bla2) inBacillus anthracisis regulated by a sigma factor (SigP) and its cognate anti-sigma factor (RsiP). Mutations leading to truncation of RsiP were previously described as a basis for PEN resistance. Here, we analyze whole-genome sequencing (WGS) data and compare the chromosomalsigP-bla1regions from 374B. anthracisstrains to determine the frequency of mutations, identify mutations associated with PEN resistance, and evaluate the usefulness of WGS for predicting PEN resistance. Few (3.5%) strains contained at least 1 of 11 different mutations insigP,rsiP, orbla1.Nine of these mutations have not been previously associated with PEN resistance. Four strains showed PEN resistance (PEN-R) by conventional broth microdilution, including 1 strain with a novel frameshift inrsiP. One strain that carries the samersiPframeshift mutation as that found previously in a PEN-R strain showed a PEN-susceptible (PEN-S) phenotype and exhibited decreasedbla1andbla2transcription. An unexpectedly small colony size, a reduced growth rate, and undetectable β-lactamase activity levels (culture supernatant and cell lysate) were observed in this PEN-S strain. Sequence analysis revealed mutations in genes associated with growth defects that may contribute to this phenotype. WhileB. anthracisrsiPmutations cannot be exclusively used to predict resistance, four of the five strains withrsiPmutations were PEN-R. Therefore, theB. anthracissigP-bla1region is a useful locus for WGS-based PEN resistance prediction, but phenotypic testing remains essential.IMPORTANCEDetermination of antimicrobial susceptibility ofB. anthracisis essential for the appropriate distribution of antimicrobial agents for postexposure prophylaxis (PEP) and treatment of anthrax. Analysis of WGS data allows for the rapid detection of mutations in antimicrobial resistance (AMR) genes in an isolate, but the presence of a mutation in an AMR gene does not always accurately predict resistance. As mutations in the anti-sigma factor RsiP have been previously associated with high-level penicillin resistance in a limited number of strains, we investigated WGS assemblies from 374 strains to determine the frequency of mutations and performed functional antimicrobial susceptibility testing. Of the five strains that contained mutations inrsiP, only four were PEN-R by functional antimicrobial susceptibility testing. We conclude that while sequence analysis of this region is useful for AMR prediction inB. anthracis, genetic analysis should not be used exclusively and phenotypic susceptibility testing remains essential.

scholarly journals Identification of veterinary pathogens by use of commercial identification systems and new trends in antimicrobial susceptibility testing of veterinary pathogens.

1994 ◽  
Vol 7 (3) ◽  
pp. 346-356 ◽  
Author(s):  
J L Watts ◽  
R J Yancey
Keyword(s):  
Antimicrobial Agent ◽  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Bacterial Species ◽  
Antimicrobial Susceptibility Testing ◽  
Human Pathogens ◽  
Host Animal ◽  
Isolation And Identification ◽  
Accurate Identification

Veterinary diagnostic microbiology is a unique specialty within microbiology. Although isolation and identification techniques are similar to those used for human pathogens, many veterinary pathogens require unique cultivation or identification procedures. Commercial identification systems provide rapid, accurate identification of human pathogens. However, the accuracy of these systems with veterinary pathogens varies widely depending on the bacterial species and the host animal from which it was isolated. Increased numbers of veterinary strains or species in the data bases of the various systems would improve their accuracy. Current procedures and interpretive criteria used for antimicrobial susceptibility testing of veterinary pathogens are based on guidelines used for human pathogens. The validity of these guidelines for use with veterinary pathogens has not been established. As with fastidious human pathogens, standardized methodologies and quality control isolates are needed for tests of organisms such as Actinobacillus pleuropneumoniae and Haemophilus somnus. Furthermore, interpretive criteria for veterinary antimicrobial agents based on the MIC for veterinary pathogens, the pharmacokinetics of the antimicrobial agent in the host animal, and in vivo efficacy of the antimicrobial agent are needed. This article reviews both the commercial identification systems evaluated with veterinary pathogens and current methods for performing and interpreting antimicrobial susceptibility tests with veterinary pathogens. Recommendations for future improvements in both areas are discussed.

scholarly journals Provisional Use of CLSI-Approved Quality Control Strains for Antimicrobial Susceptibility Testing of Mycoplasma (‘Mesomycoplasma’) hyorhinis

2021 ◽  
Vol 9 (9) ◽  
pp. 1829
Author(s):  
Lisa Käbisch ◽  
Anne-Kathrin Schink ◽  
Corinna Kehrenberg ◽  
Stefan Schwarz
Keyword(s):  
Quality Control ◽  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Antimicrobial Treatment ◽  
Antimicrobial Susceptibility Testing ◽  
Broth Microdilution ◽  
Minimal Inhibitory Concentrations ◽  
Microtiter Plates ◽  
And Staphylococcus Aureus

Antimicrobial susceptibility testing (AST) should be conducted in a standardized manner prior to the start of an antimicrobial treatment. For fastidious bacteria, such as porcine Mycoplasma (‘Mesomycoplasma’) spp., specifically M. hyorhinis, neither guidelines or standards for the performance of AST, nor quality control strains for the validation of AST results are approved by organizations like the Clinical and Laboratory Standards Institute (CLSI) or the European Committee on Antimicrobial Susceptibility Testing (EUCAST). The CLSI- and EUCAST-approved quality control strains Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 29213 were chosen to validate AST by broth microdilution using modified Friis broth, developed as growth medium for porcine Mycoplasma (‘Mesomycoplasma’) spp. The antimicrobial agents doxycycline, enrofloxacin, erythromycin, florfenicol, gentamicin, marbofloxacin, tetracycline, tiamulin, tilmicosin, tulathromycin, and tylosin were examined using customized SensititreTM microtiter plates. Minimal inhibitory concentrations, determined after 24, 48, and 72 h, were mostly within the CLSI-approved quality control ranges for defined antimicrobial agents. We propose the use of the combination of E. faecalis ATCC 29212 and S. aureus ATCC 29213 as surrogate quality control strains for the validation of future AST results obtained for M. hyorhinis by broth microdilution using modified Friis broth.

scholarly journals Antimikrobiyal Aktivitenin Belirlenmesinde Cross-Streak Metodu Kullanımı

2019 ◽  
Vol 7 (sp1) ◽  
pp. 160
Author(s):  
Mustafa Ersal
Keyword(s):  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Antimicrobial Susceptibility Testing ◽  
Rapid Screening ◽  
Resistant Bacteria ◽  
Production Environment ◽  
Specific Test ◽  
The Cross

Antimicrobial susceptibility testing can be used for prediction of therapeutic results, epidemiology and drug discovery. Microbial infections are an important problem which have developed resistance towards antimicrobial agents. Otherwise, efficacy of these agents is considerable with treatment failures associated with multidrug-resistant bacteria and it has become a global concern to public health. Therefore, explore the new antimicrobial agents and widely use of antimicrobial susceptibility need to be developed. There are many techniques for the determination of antimicrobial activity. Many of these techniques, which are applied to inhibit sensitive microorganisms, are based on diffusion-related methods in the solid or semi-solid production environment. Cross-streak among these techniques is an easy technique that allows for relatively rapid screening of cultures in research for the discovery of the new antibiotics. However, the biggest disadvantage of the Cross-streak test is the difficulty in obtaining quantitative data. Because the edges of the inhibition zone are usually very fuzzy and unclear. Some antimicrobial susceptibility testing techniques were standardized by Clinical Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) to determine the striking steps in this area. This testing procedure requires the use of specific test conditions and methods. In addition, the medium, incubation conditions and time are among these requirements. It is important to understand and develop the Cross-streak method from the currently used activity determination methods.

scholarly journals Identification, Genotyping and Antimicrobial Susceptibility Testing of Brucella spp. Isolated from Livestock in Egypt

2019 ◽  
Vol 7 (12) ◽  
pp. 603 ◽  
Author(s):  
Aman Ullah Khan ◽  
Waleed S. Shell ◽  
Falk Melzer ◽  
Ashraf E. Sayour ◽  
Eman Shawkat Ramadan ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Antimicrobial Susceptibility ◽  
Molecular Mechanisms ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Brucella Melitensis ◽  
Rpob Gene ◽  
Antimicrobial Susceptibility Testing ◽  
Animal Populations ◽  
Microbiological Methods

Brucellosis is a highly contagious zoonosis worldwide with economic and public health impacts. The aim of the present study was to identify Brucella (B.) spp. isolated from animal populations located in different districts of Egypt and to determine their antimicrobial resistance. In total, 34-suspected Brucella isolates were recovered from lymph nodes, milk, and fetal abomasal contents of infected cattle, buffaloes, sheep, and goats from nine districts in Egypt. The isolates were identified by microbiological methods and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Differentiation and genotyping were confirmed using multiplex PCR for B. abortus, Brucella melitensis, Brucella ovis, and Brucella suis (AMOS) and Bruce-ladder PCR. Antimicrobial susceptibility testing against clinically used antimicrobial agents (chloramphenicol, ciprofloxacin, erythromycin, gentamicin, imipenem, rifampicin, streptomycin, and tetracycline) was performed using E-Test. The antimicrobial resistance-associated genes and mutations in Brucella isolates were confirmed using molecular tools. In total, 29 Brucella isolates (eight B. abortus biovar 1 and 21 B. melitensis biovar 3) were identified and typed. The resistance of B. melitensis to ciprofloxacin, erythromycin, imipenem, rifampicin, and streptomycin were 76.2%, 19.0%, 76.2%, 66.7%, and 4.8%, respectively. Whereas, 25.0%, 87.5%, 25.0%, and 37.5% of B. abortus were resistant to ciprofloxacin, erythromycin, imipenem, and rifampicin, respectively. Mutations in the rpoB gene associated with rifampicin resistance were identified in all phenotypically resistant isolates. Mutations in gyrA and gyrB genes associated with ciprofloxacin resistance were identified in four phenotypically resistant isolates of B. melitensis. This is the first study highlighting the antimicrobial resistance in Brucella isolated from different animal species in Egypt. Mutations detected in genes associated with antimicrobial resistance unravel the molecular mechanisms of resistance in Brucella isolates from Egypt. The mutations in the rpoB gene in phenotypically resistant B. abortus isolates in this study were reported for the first time in Egypt.

scholarly journals Antimicrobial susceptibility testing of Neisseria gonorrhoeae and implications for epidemiology and therapy.

1993 ◽  
Vol 6 (1) ◽  
pp. 22-33 ◽  
Author(s):  
T Fekete
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
The United States ◽  
Primary Treatment ◽  
Antimicrobial Susceptibility Testing ◽  
Response To Therapy ◽  
In Vitro Tests

Antimicrobial susceptibility testing (AST) of Neisseria gonorrhoeae has been under development since the early days of antimicrobial agents. However, it is rarely applied to clinical isolates today. The history of the various in vitro tests to determine the susceptibility of N. gonorrhoeae to antibiotics is rich with evidence that these results predict response to therapy for almost all agents tested. Further, AST is a useful and important aspect of strain characterization and disease epidemiology in conjunction with the more specific but laborious techniques of auxotyping, serotyping, and plasmid analysis. Current technology has overcome many of the objections to AST for N. gonorrhoeae with standardization of test media and the development of an accurate disk diffusion AST method that is suited to most clinical laboratories regardless of volume or level of technical expertise. Ironically, the very low level of resistance to the current primary treatment strategy in the United States, ceftriaxone or another potent cephalosporin, makes the use of AST somewhat superfluous.

scholarly journals A Simplified Method for Testing Bordetella pertussisfor Resistance to Erythromycin and Other Antimicrobial Agents

2000 ◽  
Vol 38 (3) ◽  
pp. 1151-1155 ◽  
Author(s):  
Bertha C. Hill ◽  
Carolyn N. Baker ◽  
Fred C. Tenover
Keyword(s):  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Reference Method ◽  
Antimicrobial Susceptibility Testing ◽  
Disk Diffusion ◽  
Mueller Hinton ◽  
Disk Diffusion Testing ◽  
Direct Inoculation ◽  
Agar Dilution

Present methods of antimicrobial susceptibility testing ofBordetella pertussis are time consuming and require specialized media that are not commercially available. We tested 52 isolates of B. pertussis for resistance to erythromycin, trimethoprim-sulfamethoxazole, chloramphenicol, and rifampin by agar dilution with Bordet-Gengou agar (BGA) containing 20% horse blood (reference method), Etest using BGA and Regan-Lowe agar without cephalexin (RL−C), and disk diffusion using BGA and RL−C. The organisms tested included four erythromycin-resistant isolates ofB. pertussis from a single patient, a second erythromycin-resistant strain of B. pertussis from an unrelated patient in another state, and 47 nasopharyngeal surveillance isolates of B. pertussis from children in the western United States. The results of agar dilution testing using direct inoculation of the organisms suspended in Mueller-Hinton broth were within ±1 dilution of those obtained after overnight passage of the inoculum in Stainer-Scholte medium, which is the traditional method of testing B. pertussis. The Etest method produced MICs similar to those of the agar dilution reference method for three of the four antimicrobial agents tested; the trimethoprim-sulfamethoxazole results were lower with Etest, particularly when the direct suspension method was used. Most of the Etest MICs, except for that of erythromycin, were on scale. Disk diffusion testing using RL−C medium was helpful in identifying the erythromycin-resistant strains, which produced no zone of inhibition around the disk; susceptible isolates produced zones of at least 42 mm. Thus, the antimicrobial susceptibility testing of B. pertussis can be simplified by using the Etest or disk diffusion on RL−C to screen for erythromycin-resistant isolates of B. pertussis.

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