Etest (previously known as the Epsilometer test) is a way of determining antimicrobial sensitivity by placing a strip impregnated with antimicrobials onto an agar plate. A strain of bacterium or fungus will not grow near a concentration of antibiotic or antifungal if it is sensitive. For some microbial and antimicrobial combinations, the results can be used to determine a minimum inhibitory concentration (MIC). Etest is a proprietary system manufactured by bioMérieux. It is a laboratory test used in healthcare settings to help guide physicians by indicating what concentration of antimicrobial could successfully be used to treat patients' infections. [1]
Etest is a quantitative technique for determining the MIC of microoganisms. It is used for a range of Gram-negative and Gram-positive bacteria such as Pseudomonas , [2] [3] Staphylococcus , [4] and Enterococcus species, [5] as well as fastidious bacteria, such as Neisseria and Streptococcus pneumoniae. [1] It can also be used to determine MICs against certain fungi. [6] [7]
Etest is a pre-prepared non-porous plastic reagent strip with a predefined gradient of antibiotic, covering a continuous concentration range. [8] It is applied to the surface of an agar plate inoculated with the test strain, where there is release of the antimicrobial gradient from the plastic carrier to the agar to form a stable and continuous gradient beneath and in nearby to the strip.
The time taken for a plate to be ready depends on the m that is being tested, and the conditions of the agar plate.[ citation needed ] The predefined Etest gradient remains stable for at least 18 to 24 hours; that is, a period that covers the critical times of many species of fastidious and non-fastidious organisms.
After the test, the bacterial growth becomes visible after incubation and a symmetrical inhibition ellipse centered along the strip is seen. The MIC value is read from the scale in terms of μg/mL where the ellipse edge intersects the strip. After the required incubation period, the minimum inhibitory value is read where the edge of the inhibition ellipse intersects the side of the strip. The plate should not be read if the culture appears mixed or if the lawn of growth is too light or too heavy.[ citation needed ]
Etest MIC endpoints are usually clear-cut although different growth/inhibition patterns may be seen depending on the antifungal or antibiotic used. [7]
Etest can be used with many different kinds of AST agar medium as long as the medium supports good growth of the test organism and does not interfere with the activity of the antimicrobial agent. However, to maximise reproducibility, the medium chosen should fulfil the basic requirements for a susceptibility test medium. The following AST media are recommended for use with Etest:[ citation needed ]
These media may require supplemental nutrients to obtain enhanced growth of nutritionally fastidious organisms such as pneumococci, streptococci, Abiotrophia, Haemophilus, gonococci, meningococci and Campylobacter. In general, media recommendations from the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) are considered appropriate for Etest.
The Etest family of instruments is designed to simplify the daily use of Etest. Simplex C76, Nema C88, and Retro C80 are easy to use, reducing operator fatigue, saving time and improving the quality of results by increasing reproducibility. Etest and related instruments offer one of the most efficient methods for generating on-scale MIC values across 15 doubling dilutions for susceptibility testing of a wide range of drug-bug combinations, including fastidious organisms.
The Etest strip was first described in 1988 and was introduced commercially in 1991 by AB BIODISK. [5] bioMérieux acquired AB BIODISK in 2008 and continues to manufacture and market this product range under the mark Etest.
During the 1950s, Hans Ericsson (Professor of microbiology at the Karolinska Hospital and Karolinska Institute, Stockholm), the scientific founder of AB BIODISK, developed a method to standardize the disk diffusion test and to improve its reproducibility and reliability for clinical susceptibility predictions.[ citation needed ] The inhibition zone sizes from disk test results were compared to MIC values based on the reference agar dilution procedure.The correlation between zone sizes and MIC values was then assessed using regression analysis and regression lines were used for extrapolating zone interpretive limits that corresponded to the MIC breakpoint values that defined susceptible, intermediate and resistant categorical results.
Etest was first presented at the Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC) in Los Angeles in 1988 as a novel gradient concept for MIC determinations. In September 1991, Etest was launched globally as a MIC product after receiving the USA Food and Drug Administration (FDA) clearance.[ citation needed ]
An agar plate is a Petri dish that contains a growth medium solidified with agar, used to culture microorganisms. Sometimes selective compounds are added to influence growth, such as antibiotics.
A microbiological culture, or microbial culture, is a method of multiplying microbial organisms by letting them reproduce in predetermined culture medium under controlled laboratory conditions. Microbial cultures are foundational and basic diagnostic methods used as research tools in molecular biology.
Vancomycin-resistant Staphylococcus aureus (VRSA) are strains of Staphylococcus aureus that have acquired resistance to the glycopeptide antibiotic vancomycin. Bacteria can acquire resistance genes either by random mutation or through the transfer of DNA from one bacterium to another. Resistance genes interfere with the normal antibiotic function and allow bacteria to grow in the presence of the antibiotic. Resistance in VRSA is conferred by the plasmid-mediated vanA gene and operon. Although VRSA infections are uncommon, VRSA is often resistant to other types of antibiotics and a potential threat to public health because treatment options are limited. VRSA is resistant to many of the standard drugs used to treat S. aureus infections. Furthermore, resistance can be transferred from one bacterium to another.
A blood culture is a medical laboratory test used to detect bacteria or fungi in a person's blood. Under normal conditions, the blood does not contain microorganisms: their presence can indicate a bloodstream infection such as bacteremia or fungemia, which in severe cases may result in sepsis. By culturing the blood, microbes can be identified and tested for resistance to antimicrobial drugs, which allows clinicians to provide an effective treatment.
Multiple drug resistance (MDR), multidrug resistance or multiresistance is antimicrobial resistance shown by a species of microorganism to at least one antimicrobial drug in three or more antimicrobial categories. Antimicrobial categories are classifications of antimicrobial agents based on their mode of action and specific to target organisms. The MDR types most threatening to public health are MDR bacteria that resist multiple antibiotics; other types include MDR viruses, parasites.
Antibiotic sensitivity testing or antibiotic susceptibility testing is the measurement of the susceptibility of bacteria to antibiotics. It is used because bacteria may have resistance to some antibiotics. Sensitivity testing results can allow a clinician to change the choice of antibiotics from empiric therapy, which is when an antibiotic is selected based on clinical suspicion about the site of an infection and common causative bacteria, to directed therapy, in which the choice of antibiotic is based on knowledge of the organism and its sensitivities.
Burkholderia cepacia complex (BCC) is a species complex consisting of Burkholderia cepacia and at least 20 different biochemically similar species of Gram-negative bacteria. They are catalase-producing and lactose-nonfermenting. Members of BCC are opportunistic human pathogens that most often cause pneumonia in immunocompromised individuals with underlying lung disease. Patients with sickle-cell haemoglobinopathies are also at risk. The species complex also attacks young onion and tobacco plants, and displays a remarkable ability to digest oil.
Stenotrophomonas maltophilia is an aerobic, nonfermentative, Gram-negative bacterium. It is an uncommon bacterium and human infection is difficult to treat. Initially classified as Bacterium bookeri, then renamed Pseudomonas maltophilia, S. maltophilia was also grouped in the genus Xanthomonas before eventually becoming the type species of the genus Stenotrophomonas in 1993.
Burkholderia pseudomallei is a Gram-negative, bipolar, aerobic, motile rod-shaped bacterium. It is a soil-dwelling bacterium endemic in tropical and subtropical regions worldwide, particularly in Thailand and northern Australia. It was reported in 2008 that there had been an expansion of the affected regions due to significant natural disasters, and it could be found in Southern China, Hong Kong, and countries in the Americas. B. pseudomallei, amongst other pathogens, has been found in monkeys imported into the United States from Asia for laboratory use, posing a risk that the pathogen could be introduced into the country.
In microbiology, the minimum inhibitory concentration (MIC) is the lowest concentration of a chemical, usually a drug, which prevents visible in vitro growth of bacteria or fungi. MIC testing is performed in both diagnostic and drug discovery laboratories.
The disk diffusion test is a culture-based microbiology assay used in diagnostic and drug discovery laboratories. In diagnostic labs, the assay is used to determine the susceptibility of bacteria isolated from a patient's infection to clinically approved antibiotics. This allows physicians to prescribe the most appropriate antibiotic treatment. In drug discovery labs, especially bioprospecting labs, the assay is used to screen biological material and drug candidates for antibacterial activity. When bioprospecting, the assay can be performed with paired strains of bacteria to achieve dereplication and provisionally identify antibacterial mechanism of action.
Cefsulodin is a third-generation cephalosporin antibiotic that is active against Pseudomonas aeruginosa and was discovered by Takeda Pharmaceutical Company in 1977.
Mueller Hinton agar is a type of growth medium used in microbiology to culture bacterial isolates and test their susceptibility to antibiotics. This medium was first developed in 1941 by John Howard Mueller and Jane Hinton, who were microbiologists working at Harvard University. However, Mueller Hinton agar is made up of a couple of components, including beef extract, acid hydrolysate of casein, and starch, as well as agar to solidify the mixture. The composition of Mueller Hinton agar can vary depending on the manufacturer and the intended use, but the medium is generally nutrient-rich and free of inhibitors that could interfere with bacterial growth.
Mycobacteria Growth Indicator Tube (MGIT) is intended for the culture, detection and recovery of mycobacteria. The MGIT Mycobacteria Growth Indicator Tube contains 7 mL of modified Middlebrook 7H9 Broth base. The complete medium, with OADC enrichment and PANTA antibiotic mixture, is one of the most commonly used liquid media for the cultivation of mycobacteria.
Agar dilution is one of two methods used by researchers to determine the minimum inhibitory concentration (MIC) of antibiotics. It is the dilution method most frequently used to test the effectiveness of new antibiotics when a few antibiotics are tested against a large panel of different bacteria.
In microbiology, the term isolation refers to the separation of a strain from a natural, mixed population of living microbes, as present in the environment, for example in water or soil, or from living beings with skin flora, oral flora or gut flora, in order to identify the microbe(s) of interest. Historically, the laboratory techniques of isolation first developed in the field of bacteriology and parasitology, before those in virology during the 20th century.
Diagnostic microbiology is the study of microbial identification. Since the discovery of the germ theory of disease, scientists have been finding ways to harvest specific organisms. Using methods such as differential media or genome sequencing, physicians and scientists can observe novel functions in organisms for more effective and accurate diagnosis of organisms. Methods used in diagnostic microbiology are often used to take advantage of a particular difference in organisms and attain information about what species it can be identified as, which is often through a reference of previous studies. New studies provide information that others can reference so that scientists can attain a basic understanding of the organism they are examining.
Corynebacterium striatum is a bacterium that is a member of the Corynebacterium genus. It is classified as non-diphtheritic. The bacterium is a gram-positive prokaryote that assumes a 'club-like' morphology, more formally known as a corynebacteria structure. It is non-lipophilic and undergoes aerobic respiration. It is a facultative anaerobe. It is catalase negative and is an oxidase positive glucose and sucrose fermenter.
John Charles Sherris was an English-American medical doctor, pathologist, and bacteriologist. He was the president of the American Society for Microbiology (ASM) in 1983.
Schädler agar is a nutrient-rich growth medium primarily used in microbiology for the cultivation of anaerobic bacteria. It was developed to support the growth of a wide variety of anaerobic organisms, providing a conducive environment for both fastidious and non-fastidious anaerobes. The medium contains a combination of peptones, yeast extract, and other nutrients that create an optimal growth environment. Additionally, reducing agents such as cysteine and sodium thioglycolate are included to maintain the anaerobic conditions necessary for the survival of these bacteria.