The exact method used will depend largely on the type of equipment available in. Protocols used in the gfn laboratory training courses. The clinical microbiology laboratory is often a sentinel for the detection of drug resistant strains of microorganisms. The tables presented in m100 represent the most current information for drug selection, interpretation, and quality control using. The authors provide a comprehensive, uptodate procedural manual including protocols for. Gently press the discs onto the surface of the agar using flame sterilized forceps or inoculation. Nextgeneration antimicrobial susceptibility testing.
Combat antimicrobial resistance with access to the latest antimicrobials for susceptibility testing on a single platform with a system that utilizes both in vitro diagnostic ivd and research use only ruo, 96well microtitre plates for broth microdilution. Testing for these drugs is done using the kirby bauer method at this time, since they are not yet available for testing using the broth microdilution method. After the streaking is complete, allow the plate to dry for 5 minutes. Ppt antimicrobial susceptibility testing powerpoint. Use the swab with the test organism to streak a mha plate or a na plate for a lawn of growth. Influence of incubation time on antimicrobial susceptibility testing of pathogenic vibrio anguillarum and vibrio vulnificus.
Antimicrobial susceptibility testing protocols richard schwalbe, lynn steele moore, avery c. A number of antimicrobial susceptibility testing ast methods are available to determine bacterial susceptibility to antimicrobials. All of the contributors to antimicrobial susceptibility testing protocols. Kirbybauer disk diffusion susceptibility test protocol, inoculation of the muellerhinton agar plate. The use of these plates, with a documented test protocol, including specification of. Kirbybauer disk diffusion susceptibility test protocol asm. Approved standard 9th edition, represents the standard for clinical laboratories performing susceptibility testing today. Disc diffusion techniques are used by most laboratories to test routinely for antimicrobial susceptibility. Because susceptibility can vary even within a species with some strains being more resistant than others, antibiotic susceptibility testing ast is usually carried out to determine which antibiotic will be most successful in treating a bacterial infection in vivo. Antimicrobial susceptibility testing protocols respectfully and lovingly present our work as a token of our admiration to rick and in the hope that through our efforts we may carry on his passion and love of susceptibility testing and microbiology. Antimicrobial susceptibility testing agar dilution. You have full access to read online and download this title. Change of antibiotic susceptibility testing guidelines from clsi to eucast. The protocols and parameters set by clsi are considered to be the gold standard in the united states and are used by regulatory authorities, such as the fda, to make evaluations.
It covers stepbystep protocols with an emphasis on optimizing the detection of resistant microorganisms. Manual for the laboratory identification and antimicrobial. There are few activities in the clinical microbiology laboratory that utilize more technologist time and laboratory resources than antimicrobial susceptibility testing ast. Who headquarters, lyon office, regional offices, national offices, and collaborating centres are active in supporting training activities, quality assurance programs, and resources to laboratories. Validation of an antimicrobial susceptibility testing. Surprisingly, one of these is the disk diffusion method first published in 1966 and the various iterations thereof. Clsi subcommittee for antimicrobial susceptibility testing ast subcommittee revised the enterobacteriaceae breakpoints. About clsi antimicrobial and antifungal susceptibility testing resources. Antimicrobial susceptibility testing protocols 1st edition richard. A general method for rapid determination of antibiotic. Antimicrobial and antifungal susceptibility testing resources. Purpose the purpose of the kirbybauer disk diffusion susceptibility test is to determine the sensitivity or resistance of pathogenic aerobic and. Determination of minimum inhibitory concentrations mics.
Of particular importance is the concept of efficacy ratios. Clsi develops standardized reference methods that measure the susceptibility of bacteria and fungi to antimicrobial agents in vitro. Validation of an antimicrobial susceptibility testing protocol for brachyspira hyodysenteriae and brachyspira pilosicoli in an international ring trial. From a microbiological and clinical perspective, the science and medicine of antimalarial susceptibility testing lag far behind those for bacteria, fungi, and viruses. With over 300 antimicrobials available in extended dilution ranges, the sensititre system allows you to consolidate confirmatory tests. Antimicrobial susceptibility testing journal of clinical.
Antimicrobial susceptibility testing protocols ebook. Pdf a method for antibiotic susceptibility testing. Mic methods are widely used in the comparative testing of new agents. The second section describes specialized susceptibility protocols such as surveillance procedures for detection of antibiotic resistant bacteria, serum bactericidal assays, timekill curves, population analysis, and synergy testing. Antimicrobial susceptibility testing microbiology resource center. There may be some inhibition of growth and the organism. Synergy screen test synergy with high level aminoglycoside gentamicin 500 ugml.
Gives laboratory personnel an integrated resource for labbased techniques and charts within the contextual role of clinical microbiology in modern medicine. Antimicrobial susceptibility testing thermo fisher. Antimicrobial susceptibility testing protocols richard schwalbe, lynn steelemoore, avery c. Manual for the laboratory identification and antimicrobial susceptibility testing of bacterial pathogens of public health importance in the developing world.
To ensure correct antibiotic treatment and reduce the unnecessary use of antibiotics, there is an urgent need for new rapid methods for species identification and determination of antibiotic susceptibility in infectious pathogenic bacteria. Antimicrobial susceptibility testing protocols crc press. Most laboratories now download their data from their laboratory information management. Antimicrobial susceptibility testing protocols clearly defines the role of the clinical microbiology laboratory in integrated patient care and provides a comprehensive, uptodate procedural manual that can be used by a wide variety of laboratorians. For antimicrobial susceptibility testing of escherichia. A limited number of methods for antimicrobial susceptibility testing ast of medically important microorganisms have survived the maturation of modern diagnostic clinical microbiology. This document includes updated tables for the clinical and laboratory standards institute antimicrobial susceptibility testing standards m02, m07, and m11. Antimicrobial susceptibility testing ast types and. Susceptibility testing of medically important parasites. Antimicrobial susceptibility testing protocols 1st. Haemophilus influenzae, neisseria meningitidis, streptococcus pneumoniae, neisseria gonorrhoeae, salmonella serotype typhi, shigella, and vibrio cholerae. A free powerpoint ppt presentation displayed as a flash slide show on id. Change of antibiotic susceptibility testing guidelines.
Clsi, eucast, and others describe strategies and protocols for assuring the quality of antimicrobial susceptibility test results. Kirbybauer disk diffusion susceptibility test protocol. Antimicrobial susceptibility testing ast results have a significant impact when managing patients with serious infections. Additional protocols related to antimicrobial resistance, e. The standard protocol must be followed exactly for you, or any clinical lab, to interpret the results reliably. Verification of antimicrobial susceptibility testing. Dilution methods are used to determine the minimum inhibitory concentrations mics of antimicrobial agents and are the reference methods for antimicrobial susceptibility testing against which other methods, such as disk diffusion, are calibrated. Antimicrobial susceptibility testing protocols richard. Non fdaapproved or modified fdaapproved or cleared test verification of ast systems has gotten a lot of attention. The aim of broth and agar dilution methods is to determine the lowest concentration of the assayed antimicrobial agent minimal inhibitory concentration, mic that, under defined test. On three independent occasions, the isolates were cultured from frozen stock at each laboratory, subcultured twice after thawing on faba with 10 % horse or sheep blood faba, and tested for antimicrobial susceptibility. Nowadays, the mic is used in antimicrobial susceptibility testing. Updated protocol for antimicrobial susceptibility testing. A number of antimicrobial susceptibility testing ast methods are available.
The protocol was approved by the veterinary antimicrobial susceptibility testing vast subcommittee of the clsi in june 2015 and then applied to a collection of 65 german a. Inoculate the plate with the test organism by streaking the swab in a backandforth motion very close together as you move across and down the plate. Updated protocol for antimicrobial susceptibility testing of. The national antimicrobial resistance monitoring system. The national antimicrobial resistance monitoring system narms, established in. It has been suggested that, at least in terms of direct relevance to the care of patients with infection, ast may be the. Below you find protocols that have been used in the gfn laboratory training courses. Antimicrobial susceptibility testing of arcobacter. The clsi susceptibility testing document lists groups of some antimicrobial agents with nearly identical activities that can provide practical alternatives for testing. M100 performance standards for antimicrobial susceptibility testing, 30th edition. A disc of blotting paper is impregnated with a known volume and appropriate concentration of an antimicrobial, and this is placed on a plate of susceptibility testing agar uniformly inoculated with the test. We have developed a general method for the rapid identification of the bacterial species causing an infection and the determination of their antibiotic. The selection of a method is based on many factors such as practicality. Antibiotic discs can be placed on the surface of the agar using sterilized forceps.
Recent developments in identification tools can now identify microbes with unprecedented speed, but phenotypic susceptibility testing has lagged behind. Performance standards for antimicrobial susceptibility testing. In clinical laboratories they are used to establish the susceptibility of. In this document, we attempt to answer many of the frequently asked questions regarding antimicrobial susceptibility testing.