DIFFERENTIAL CULTURE MEDIUM CONTAINING DUAL CARBAPENEM

Abstract

This invention relates to the fields of microbiology and clinical diagnosis, and specifically to a differential medium containing dual-carbapenems that enables the selective detection of carbapenem resistance in Gram-negative bacteria. The invention allows for the isolation of carbapenem-resistant strains of Enterobacterales and non-fermenting Gram-negative bacteria, distinguishing them from susceptible strains.

Description

DIFFERENTIAL CULTURE MEDIUM CONTAINING DUAL CARBAPENEM Technical Area This invention relates to the field of microbiology and clinical diagnosis, particularly in Gram-negative bacteria. Dual-carbapenem containing, enabling selective detection of carbapenem resistance. The invention relates to a differential culture medium. The invention concerns Enterobacterales and non-fermenting Gram-negative bacteria. isolating carbapenem-resistant strains of bacteria from susceptible strains provides. Previous Technique Carbapenem-resistant Gram-negative bacteria are particularly common in intensive care units. Hospital-acquired infections lead to significant mortality and morbidity. Clinical early and accurate detection of these resistant microorganisms in microbiology laboratories Detection is critical for infection control and determining appropriate treatment strategies. It is important. Most current selective media contain a single carbapenem antibiotic. It has the following technical limitations: • Because they contain only one carbapenem, they are resistant to all resistance mechanisms. their inability to provide sufficient selectivity, • Non-fermenting Gram-negative bacteria (e.g., Pseudomonas aeruginosa, (Acinetobacter spp.) insufficient suppression or leading to false negative results opening • Distinguishing morphological features of differential culture media by antibiotic addition deterioration, • In clinical samples containing dense polymicrobial flora (such as perirectal or groin swabs) Low density inability to detect resistant strains. For these reasons, both Enterobacterales and non-fermenting Gram-negative bacteria A new generation of systems capable of simultaneously and selectively and reliably detecting carbapenem-resistant strains. Culture medium is required. 1 Brief Description of the Invention The main purpose of this invention is to eliminate the problems mentioned above by developing Carbapenems. Selective development of resistant Gram-negative bacteria, carbapenem-sensitive strains suppression, preservation of differential colony morphology, and Enterobacterales and non- an improved culture medium that enables reliable separation of fermenters in the same environment It is the development of. The invention describes the use of an Eosin Methylene Blue (EMB) based differential culture medium to culture two different carbapenems. It is based on the principle of using the antibiotics ertapenem and meropenem in combination. This dual- Thanks to the carbapenem combination, carbapenem-resistant Enterobacterales and non- The growth of fermentative Gram-negative bacteria is made possible. In addition, The growth of carbapenem-sensitive strains is effectively suppressed, and EMB medium distinguishes them. The distinctive differential properties (e.g., metallic luster for E. coli) are preserved. The invention has clinical implications. For early screening and isolation purposes in samples, high selectivity and sensitivity. It has been improved. Detailed description of the invention The subject of this invention is to achieve dual-carbapenem selectivity using ertapenem and meropenem. Differential culture media will be developed by using antibiotics together in the same culture medium. This This allows bacteria with different carbapenemase types and resistance mechanisms to work together in the same way. This will ensure that it can be selected in the environment. Ertapenem is particularly effective against Enterobacterales. While increasing selectivity, meropenem inhibits resistant strains of non-fermenting Gram-negative bacteria. This supports the finding. EMB-based medium for preserving differential characteristics, lactose fermentation, and dyeing. This will allow the observation of different colony morphologies through interaction. Dual- Even with carbapenem supplementation, resistant Escherichia coli strains develop metallic, shiny colonies. Its morphology will be preserved. In addition, other Enterobacterales species have characteristic color and colony structure. This will continue. This facilitates diagnostic interpretation and avoids false positive / negative results. It will reduce the risk. The culture medium developed within the scope of this invention is suitable for samples containing dense flora, such as perirectal and inguinal swabs. selective carbapenem-resistant bacteria, which are found in low densities even in clinical samples. This makes it possible to determine this. 2 In the preferred application of the invention, 0.01 mg of 1 gram of lyophilized Ertapenem antibiotic powder is obtained. (milligram) and 0.01 mg (milligram) of 1 g of Meropenem lyophilized powder antibiotic are weighed. in this hand The resulting dual antibiotic mixture is diluted with at least 10 times its weight in water. Preferably 5-30 ml. Diluted dual antibiotics are added to Eosin Methylene blue agar (EMB) weighing 1 / 2. The prepared agar can be stored in the refrigerator at 2-8°C for 15 days before use. Every 15 days... Since the effectiveness of antibiotics will decrease after a few days, they will need to be renewed. The invention... In another application, dual will be stored in its powder form before dilution. The antibiotic powder mixture is diluted and mixed with agar before use for testing. It will be ready immediately. Industrial and Clinical Applications The invention may be used in the following areas: • Infection control screenings in intensive care units, • Early detection of carbapenem-resistant colonization, • Rapid isolation of clinical swab and urine samples, • Diagnosis of resistant Enterobacterales and non-fermenters, • Outbreak research and CRO studies, • All clinical applications requiring routine and rapid screening cultures. Example Applications 1 Testing with Reference Strains Carbapenem-resistant and susceptible ATCC reference strains were cultured in the culture medium covered by this invention. They were cultured comparatively. Resistant strains developed, while susceptible strains remained. It was observed that the colony morphology was suppressed; the EMB-specific morphology was preserved. Example Applications 2 Clinical Urine Sample The carbapenem-resistant Pseudomonas strain obtained from a clinical urine sample is the subject of the invention. It was isolated as a single colony on the culture medium. Example Applications 3 – Perirectal Swab In perirectal swab samples containing dense flora, carbapenem-resistant bacteria are found in low numbers. It was selectively detected even at high concentrations. 3

Claims

1. Selective detection of carbapenem resistance in Gram-negative bacteria. It is a differential culture medium; an Eosin Methylene Blue (EMB) based differential medium. culture medium matrix, ertapenem as the first carbapenem antibiotic, second carbapenem Characterized by containing meropenem as an antibiotic and being carbapenem-resistant. Allows the growth of Enterobacterales and / or non-fermenting Gram-negative bacteria. Differential agar suppresses the growth of carbapenem-sensitive strains when administered.

2. According to Claim 1, the differential medium in question is the ertapenem and meropenem, Simultaneous selection of Gram-negative bacteria with different carbapenemase types characterized by its presence in the environment within the concentration ranges that will provide differential culture medium.

3. Differential culture medium according to claim 1 or 2, for Enterobacterales species. while allowing colony development of carbapenem-resistant strains of the same species Characterized by suppressing the growth of carbapenem-sensitive strains. differential culture medium.

4. Differential medium according to any of claims 1–3, non-fermenting Gram-negative. negative bacteria, especially Pseudomonas aeruginosa and / or Acinetobacter by enabling the selective development of carbapenem-resistant strains of species Differential culture medium that has been characterized.

5. Differential medium according to any of claims 1–4, EMB-based differential. the distinctive colony morphology of the medium, dual-carbapenem antibiotic content Differential agar characterized by its preservation properties.

6. Differential medium according to claim 5, for carbapenem-resistant Escherichia coli. characterized by the metallic sheen colony morphology of the strains. differential culture medium obtained.

7. Differential culture medium according to any of claims 1–6, and highly polymicrobial. Carbapenem-resistant bacteria found in low concentrations in clinical samples containing flora. characterized by its ability to allow selective detection of bacteria. differential culture medium.

8. Differential culture medium according to any of claims 1–7; intensive care infection in control screenings, in the detection of carbapenem-resistant colonization, clinical swab 4 and / or selective carbapenem-resistant Gram-negative bacteria in urine samples Used for insulation purposes.

9. Use according to Claim 8, and the clinical samples in question are perirectal swabs, groin. The use is characterized by having a swab or urine sample.