KR102511614B1 - Reagent Composition, Kit, and Method for the Detection of Gram-Negative Bacteria Acinetobacter Baumannii - Google Patents

Abstract

상기 화학식 1에서,
R¹ 및 R²는, 각각 독립적으로 탄소수 1 내지 5개의 직쇄 또는 분지쇄 알킬이다.The present invention relates to a reagent composition for detecting the Gram-negative bacteria Acinetobactger baumanii , a kit for diagnosing Acinetobactger baumani infection, and a method for detecting Acinetobactger baumanii. According to the present invention, only Acinetobacto baumani can be selectively detected by using the compound represented by Formula 1 below:
[Formula 1]

In Formula 1,
R ¹ and R ² are each independently a straight-chain or branched-chain alkyl having 1 to 5 carbon atoms.

Description

Reagent Composition, Kit, and Method for the Detection of Gram-Negative Bacteria Acinetobacter Baumannii}

The present invention relates to a reagent composition, a kit, and a detection method for detecting Gram-Negative bacteria Acinetobacter Baumannii .

Worldwide, pathogens exhibiting multi-drug resistance are greatly increasing due to the misuse and abuse of antibiotics, and the growing threat of bacteria and drug resistance continues to harm humans. Accordingly, many organizations, such as the Centers for Disease Control and Prevention (CDC) and the World Health Organization (WHO), have reported that the emergence and spread of multidrug-resistant bacteria has led to global problems such as increased mortality, prolonged treatment period, and increased medical costs. recognized as a serious threat to public health. Accordingly, in 2017, the World Health Organization identified six bacteria that pose the greatest threat to human health [ESKAPE: Enterococcus faecium (EF), Staphylococcus aureus (SA), Klebsiella pneumoniae (KP), Acinetobacter baumannii (AB), Pseudomonas aeruginosa (PA) and Enterobacter species (ES)].

Identification of bacterial strains is very important for the diagnosis, prevention and treatment of infectious diseases caused by bacteria. To diagnose and identify these bacteria, bacterial culture is required, and differential staining to identify them is generally performed. However, common staining methods require several steps, and misdiagnosis may occur depending on the staining method and reading. Recently, fluorescent staining based on dyes or target molecules has attracted attention as an alternative to these traditional differential staining techniques. This fluorescence imaging technology can practically quantify bacteria through real-time monitoring (survival/death), and is usefully used in various clinical fields due to the high selectivity and sensitivity of fluorescence. Accordingly, it has been recognized that there is a need for improved methods for imaging specific bacteria. In particular, the development of a fluorescence-based staining technique for clearly identifying and diagnosing pathogens such as ESKAPE is highly needed.

One object of the present invention is to provide a reagent composition capable of selectively detecting specific bacteria.

Another object of the present invention is to provide a diagnostic kit capable of selectively detecting a specific bacterium.

Another object of the present invention is to provide a method for selectively detecting specific bacteria.

In the present invention, a bacteria-dye combination screening (BDCS) approach, which is a new approach for identifying and diagnosing biofilms caused by target bacteria that may occur in medical devices (catheters, etc.) as well as specific bacteria, is newly introduced. introduced and used. In the present invention, among the ESKAPE strains in the BDCS development process, drug-sensitive (particularly, carbapenem-sensitive) Acinetobacter baumannii (carbapenem-sensitive Acinetobacter baumannii ) and drug-resistant (particularly, carbapenem-resistant) Acinetobacter baumanni (CRPA) , carbapenem-sensitive A. baumannii ) was confirmed as a dye capable of selective fluorescence imaging, as well as it was confirmed that Acinetobacter baumani bacteria and biofilm selectively capable of fluorescence imaging in a urinary catheter.

Accordingly, according to one aspect of the present invention, a reagent composition for selectively detecting Acinetobacter baumannii containing a compound represented by Formula 1 is provided:

[Formula 1]

In Formula 1, R ¹ and R ² are each independently a straight-chain or branched-chain alkyl having 1 to 5 carbon atoms.

According to another aspect of the present invention, a kit for diagnosing infection of Acinetobacter baumannii is provided, including the compound represented by Formula 1 above.

According to another aspect of the present invention, a method for selectively detecting Acinetobacter baumannii is provided, comprising the step of bringing the compound represented by Formula 1 into contact with an analyte.

According to one embodiment of the present invention, the selective detection method may further include observing fluorescence of the analyte.

Since the compound according to the present invention selectively fluoresces only Acinetobacto baumannii, it is possible to selectively detect only Acinetobacto baumannii among various bacteria. Accordingly, it is possible to selectively, efficiently, simply, and quickly diagnose and monitor only Acinetobacto baumannii by using the reagent composition, diagnostic kit, and detection method containing the compound of the present invention. In particular, Acinetobacter baumani is a type of superbacteria that is a major cause of urinary tract infections, so it can be used to select bacteria that cause urinary tract infections or to monitor contamination of medical instruments in real time.

1 is a diagram showing the results of bacterial-dye combination screening using BMeS-pA, coumarin 120, and rhodamine 123 for multidrug-resistant gram-negative and positive bacteria.
2 is a graph showing changes in bacterial cell membrane zeta potential by BMeS-pA (the compound of Formula 2).
3 is a graph showing absorption and fluorescence spectra of BMeS-pA.
4 is a graph showing the stability test results of BMeS-pA under various conditions such as pH buffer, temperature, light, serum and biometal.
5 is a graph showing the results of the minimum inhibitory concentration test for confirming the antibacterial effect of BMeS-pA.
6 is a diagram showing the results of bacterial biofilm fluorescence imaging by BMeS-pA.
7 is a picture showing an image showing fluorescence by the compound of the present invention in a urinary catheter coated with a biofilm of a specific bacteria.

Hereinafter, the present invention will be described in detail.

The terms used in this application are only used to describe specific embodiments and are not intended to limit the present invention. Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs.

Throughout the specification, when a part “includes,” “includes,” or “has” a certain component, it means that it may further include other components unless otherwise specifically defined.

The present invention relates to a reagent composition for selective detection of Acinetobacter baumannii , a kit for diagnosing infection of Acinetobacter baumannii, and Acinetobacter baumannii, characterized in that it contains a compound represented by Formula 1 below. A selective detection method for Bacto baumannii ( Acinetobacter baumannii ) is provided:

[Formula 1]

In Formula 1, R ¹ and R ² are each independently a straight-chain or branched-chain alkyl having 1 to 5 carbon atoms, and specifically, each independently may be a straight-chain or branched-chain alkyl having 1 to 3 carbon atoms.

The compound of Formula 1 may be specifically represented by Formula 2 below:

[Formula 2]

The compound of Formula 1 is a single benzene ring-based fluorescent probe. Benzene is the simplest aromatic hydrocarbon with six carbon rings and is one of the most basic structural units for constructing pi-conjugation systems, which are widely used as fluorescent dyes for fluorescence imaging, displays, and other luminescent materials due to their enhanced spectral signal. The compound of Formula 1 of the present invention exhibits high fluorescence in liquid and solid states, and is chemically stable under light irradiation and various acidity conditions. The compound of Chemical Formula 1 is a positive charge green fluorescent probe containing two amines (—NH ₂ ), and exhibits light absorption at 377 nm and fluorescence at 517 nm.

In an embodiment of the present invention, fluorescence imaging screening was performed on 8 Gram-negative bacteria and 4 Gram-positive bacteria (including multidrug-resistant bacteria) using the fluorescent probe, that is, the compound of Formula 1 (FIG. 1). Through this, it was confirmed that the compound of Formula 1 of the present invention selectively fluorescently stains the Gram-negative bacteria Acinetobacto baumani sensitive and resistant strains (FIG. 1).

Commercially available bacterial fluorescence staining probes can be applied to fluorescence imaging by largely distinguishing between Gram-negative bacteria and Gram-positive bacteria, but a fluorescence probe capable of fluorescence imaging targeting a specific bacteria has not been reported to date. The compound of Formula 1 of the present invention can selectively perform fluorescence imaging on specific bacteria (Gram-negative Acinetobacter baumani susceptible and resistant strains), and can be used in reagent compositions, kits, etc. to selectively detect only the bacteria. there is.

Currently, as fluorescent probes for staining bacteria, SYTO 9, PI (Propidium Iodine), Alexa-Fluoro, etc. are commercially available. However, these fluorescent probes are multi-aromatic-based phosphors and have problems such as low solubility in most organic solvents and low photostability due to their large molecular weight and tight bonds. The compound represented by Chemical Formula 1 of the present invention has high photostability, thermal stability, stability in serum and compatibility with acidity in vivo due to the nature of the material. Therefore, the compound of the present invention can be applied not only for biological research, but also for bacterial fluorescence imaging of high resolution (super-resolution microscopy) through these characteristics.

In this regard, in an embodiment of the present invention, the zeta-potential of the cell membrane of Acinetobacter baumani (susceptible/resistant strain) was investigated by the compound of Formula 1, and the compound of the present invention When stained with, it was confirmed that the zeta potential of the Acinetobacter baumani strain cell membrane increased (FIG. 2). At this time, Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA, methicillin-resistance S. aureus ) were representative of Gram-positive bacteria not stained with BMeS-pA and were used as controls. In an embodiment of the present invention, the optical properties of BMeS-pA were analyzed based on the above results, and as a result, bio-metal; It was confirmed that there was no specific reaction to FeCl ₂ , FeCl ₃ , CuCl ₂ , CaCl ₂ , ZnCl ₂ , KCl, NaCl ₂ , MgCl ₂ , pH 7.4 buffer, light, temperature (50 ℃) and serum albumin ( bovine serum albumin, BSA) was confirmed to be stable (Figs. 3-4).

According to one embodiment of the present invention, the Acinetobacto baumani selectively detected in the reagent composition, infection diagnosis kit, and detection method of the present invention is specifically multi-drug resistant Acinetobacto baumani , More specifically, it may be carbapenem-resistant Acinetobacto baumani.

The Acinetobacter baumani can generate a biofilm on a medical device. In particular, the Acinetobacto baumani may contaminate the urinary catheter and form a biofilm on the urinary catheter to exist. infection can occur. In an embodiment of the present invention, a biofilm formed by specific bacteria (AB/CRAB, SA/MRSA) on the surface of a urinary catheter, which is a medical device, was subjected to fluorescence imaging using the compound of the present invention. It was confirmed by confocal laser scanning microscopy (CLSM) that the biofilm formed by B. baumani was fluorescently stained with the compound of the present invention. On the other hand, it was confirmed that the biofilm formed by Staphylococcus aureus on the catheter surface was not fluorescently stained (FIG. 6-7). Therefore, according to one embodiment of the present invention, the reagent composition of the present invention can be used to selectively detect a biofilm produced by Acinetobacto baumani, wherein the biofilm is specifically a medical device, more specifically may be formed in the urinary catheter. According to another embodiment of the present invention, the selective detection method of Acinetobacto baumani of the present invention comprises the step of contacting the compound represented by Formula 1 with a medical device, more specifically, a urinary catheter as an analysis object. can

Hereinafter, a preferred embodiment is presented to aid understanding of the present invention. However, the following examples are provided to more easily understand the present invention, and the content of the present invention is not limited by the following examples.

Example 1. Bacterial fluorescence imaging screening

Fluorescence imaging screening was performed on Gram-negative bacteria and Gram-positive bacteria, including multidrug-resistant bacteria, using the compounds of Formulas 2, 3, or 4 below, and the results are shown in FIG. 1 .

[Formula 2]

[Formula 3]

[Formula 4]

The compound of Formula 2 is a compound according to the present invention.

The compound of Formula 3 represents coumarin 120 (7-amino-4-methylcoumarin, coumarin 120), which is a fluorescent dye. Coumarin 120 has a molecular weight of 175.2 g/L, and exhibits absorption at 345 nm and fluorescence at 445 nm.

The compound of Formula 4 represents rhodamine 123 among fluorescent dyes. Rhodamine 123 has a molecular weight of 287.33 g/L, absorbs light at a wavelength of 512 nm and emits fluorescence at 531 nm.

In the present invention, representatively selected Gram-negative bacteria, Acinetobacter baumani sensitive and resistant strains and Gram-positive bacteria, Staphylococcus aureus sensitive and resistant strains, the fluorescence imaging characteristics of the compounds of Formulas 2, 3, and 4 were observed. Information on Gram-negative bacteria and Gram-positive bacteria, including multidrug-resistant bacteria for screening bacteria-dye combinations, is specified in Table 1 below.

[Table 1]

Specifically, strains for bacterial-dye combination screening were inoculated into CA-MHB (Ca ²⁺ Muller Hilton broth) medium and then cultured at 37° C. for 24 hours. After overnight culture, centrifuge the bacterial culture medium at 13,000 rpm for 5 minutes using a 1.5 mL microtube and discard the supernatant. The bacterial pellet remaining at the bottom of the microtube is washed three times with phosphate buffered saline (PBS, pH 7.4). After washing, the remaining bacterial pellet is resuspended in phosphate buffered saline to adjust to about 1 x 10 ⁸ CFU/mL. A dye for screening, BMeS-pA, coumarin 120, and rhodamine 123 were added to the bacterial suspension at 100 μM, 100 μM, and 15 μM, respectively, and then incubated at 37° C. for 1 hour. After incubation for 1 hour, after centrifugation at 13,000 rpm for 5 minutes, discard the supernatant and wash thoroughly with phosphate buffered saline (pH 7.4) three times to remove the remaining dye. Fluorescent images of the stained bacteria were observed through CLSM, and for this, the washed bacterial pellet was resuspended in phosphate buffered saline and spread on a slide glass. Fluorescence imaging of bacteria of each dye is 405 nm excitation / 519 nm emission for BMeS-pA (compound of formula 2), 405 nm excitation / 460 nm emission for coumarin 120 (compound of formula 3) and rhodamine 123 (formula 4 compound) was performed under conditions of 488 nm excitation/560 nm emission.

As a result of the bacterial-dye combination screening for Acinetobacter baumani susceptible and resistant strains and Staphylococcus aureus susceptible and resistant strains, in the case of Coumarin 120 (compound of Formula 3), fluorescence imaging was not performed in Gram-negative and Gram-positive bacteria, Rhoda In the case of Min (compound of Formula 4), it was confirmed that staining was well performed in both Gram-negative and Gram-positive bacteria without selectivity. On the other hand, in the case of BMeS-p-A (the compound of Formula 2), it was confirmed that the gram-negative Acinetobacter baumani sensitive and resistant strains were selectively fluorescently stained. In addition, as a result of fluorescence intensity analysis, it was confirmed that BMeS-p-A selectively increased fluorescence only in Acinetobacter baumani sensitive and resistant strains (Fig. 1a, b).

Based on this, in order to further clarify the characteristics of bacterial-selective fluorescence imaging of BMeS-p-A, additional bacterial-dye combination screening for 6 Gram-negative bacteria and 2 Gram-positive bacteria was performed in the same manner as above.

As a result, BMeS-p-A confirmed that fluorescence imaging was impossible except for Acinetobacter baumani susceptible and resistant strains of Gram-negative bacteria (FIG. 1c).

Example 2. Observation of zeta-potential change of bacterial cell membrane by BMeS-p-A

According to the bacterial-dye combination screening results, it was decided to select BMeS-p-A capable of fluorescent staining of a specific bacterium, and to investigate the interaction between BMeS-p-A and the bacterial cell membrane, the zeta potential of the bacterial cell membrane was measured. Results are shown in FIG. 2 .

BMeS-p-A is a positively charged substance with two amine groups, and is expected to have high binding affinity to bacterial cell membranes with a relatively negative charge. Therefore, the zeta potential of the bacteria was measured to observe the change in the potential difference of the bacterial cell membrane according to the BMeS-p-A treatment. In this experiment, Acinetobacter baumani sensitive and resistant strains that showed fluorescence selectively to BMeS-p-A were used, and Gram-positive Staphylococcus aureus sensitive and resistant strains that did not show any fluorescence were used as a control.

Looking more specifically, as in Example 1, the four strains were inoculated into CA-MHB medium and then cultured at 37° C. for 24 hours. After overnight incubation, the bacterial culture is centrifuged at 13,000 rpm for 5 minutes, and the supernatant is discarded. Bacterial residues from which the supernatant has been removed are washed three times with phosphate buffered saline (pH 7.4), and then resuspended in phosphate buffered saline to adjust to about 1 x 10 ⁸ CFU/mL. After treating the bacterial suspension with 100 µM of BMeS-pA, it was incubated at 37°C for 1 hour. After incubation for 1 hour, the 4 kinds of bacterial cultures stained were once again centrifuged to remove the supernatant, and washed three times with phosphate buffered saline to remove the remaining BMeS-pA. After the washed bacteria were resuspended in sterile water, the charge on the bacterial surface was evaluated by measuring the zeta potential using a Zetasizer Nano ZS90 (Malvern Instruments).

As a result, the Acinetobacter baumani susceptible and resistant strains not treated with BMeS-p-A measured -45.2 mV and -46.9 mV, respectively, and the Staphylococcus aureus susceptible and resistant strains showed -39.2 mV and -35.5 mV, respectively. was (FIG. 2). As a result of measuring the zeta potential of bacteria after treatment with BMeS-p-A, Acinetobacter baumani sensitive and resistant strains showed increased potential compared to the group not treated with BMeS-p-A. In contrast, no significant potential change was observed in the strains susceptible and resistant to Staphylococcus aureus compared to the group not treated with BMeS-p-A.

These results show that Gram-negative bacteria have a slightly higher negative charge than Gram-positive bacteria because they contain lipopolysaccharide (LPS) in their outer membrane. Due to the characteristics of these gram-negative bacteria, it is believed that it binds to BMeS-p-A at a higher level. Therefore, it is considered that BMeS-p-A can be used as a fluorescent dye suitable for selective fluorescence imaging. In addition, the positive charge of coumarin 120 containing one amine used is not sufficient to label the bacterial surface, and rhodamine 123 containing two cationic amines does not seem to be suitable in terms of selectivity.

Example 3. Confirmation of Absorption/Fluorescence Characteristics of BMeS-p-A

In Example 1, absorption and fluorescence graphs were obtained for BMeS-p-A, which can selectively perform fluorescence imaging of Acinetobacter baumani sensitive and resistant strains, and the results are shown in FIG. 3.

BMeS-p-A was measured using a spectrophotometer (UV/Vis spectrophotometer; Agilent Technologies Cary 8454, US) and a fluorometer (spectro-fluorophotometer; SHIMADZU CORP. RF-6000, Japan) to examine the absorption and emission spectra (UV/Vis spectrophotometer) of BMeS-p-A. Vis absorption and emission spectra) were obtained, and a standard quartz cuvette (Hellma Analytics, Jena, Germany) was used for absorption and fluorescence measurement. The indicated graph is the result of absorption and emission fluorescence graphs of BMeS-p-A in the solvent of pH 7.4 phosphate buffered saline. Under that condition, BMeS-p-A showed maximum absorbance at 375 nm and maximum green fluorescence at 515 nm ( Fig. 3).

Example 4. Confirmation of stability of BMeS-p-A under various conditions

Considering the possibility of further practical application of BMeS-p-A, which showed the specific bacterial fluorescence imaging results in Example 1, the stability of BMeS-p-A was confirmed under various conditions, and the results are shown in FIG. 4a.

pH 7.4 buffer (culture medium condition), temperature of 50°C (thermal stability), bovine serum albumin (BSA; 35 mg/mL, this concentration is based on human blood serum concentration) and light stability. The stability of BMeS-p-A was evaluated under the condition of UV light (365 nm, 3W) for light. Specifically, after adding BMeS-p-A (10 μM) to each condition and reacting at room temperature for 60 minutes, the change in fluorescence spectrum was confirmed. The graph indicated in FIG. 4a compares the value of 515 nm showing the maximum fluorescence in the fluorescence spectrum obtained at the beginning and after 1 hour in each condition.

As a result, fluorescence change due to pH7.4 buffer, light irradiation, and bovine serum albumin was not observed, and a slight decrease in fluorescence intensity was observed by light irradiation, but this was considered a negligible fluorescence change as not significant.

In addition, fluorescence intensity change after reacting BMeS-pA with metal ions to evaluate the stability of FeCl ₂ , FeCl ₃ , CuCl ₂ , CaCl ₂ , ZnCl ₂ , KCl, NaCl ₂ , MgCl _{2 ,} which are metal ions present in vivo. was observed, and the results are shown in Figure 4b.

As a result, no fluorescence change of BMeS-p-A by the biometal was observed.

Based on these results, it was confirmed again that stable fluorescence characteristics can be exhibited without external interference when conducting fluorescence imaging studies of bacteria.

Example 5. Minimum inhibitory concentration test to confirm the antibacterial effect of BMeS-p-A

The antibacterial effect of BMeS-p-A against 12 types of bacteria, including multi-resistant bacteria, was confirmed, as shown in FIG. 5.

In order to measure the antibacterial activity against 12 types of bacteria, including multidrug-resistant bacteria, the Minimum Inhibitory Concentration (MIC) of bacteria was investigated according to the CLSI (clinical and laboratory standard institute, 2016) guidelines. A broth micro-dilution method using a 96-well plate was used, and the same CA-MHB as used in Example 1 was used as the medium. The six standard strains used in this experiment were distributed from the strain bank ATCC (American type culture colletion) and CCARM (culture collection of antibiotics resistant microbes), and the six resistant strains were distributed from Asan Medical Center (AMC). received, and information on a total of 12 strains is shown in Table 1.

Specifically, 12 strains, including multidrug-resistant bacteria, were cultured on a blood agar plate at 37° C. for 24 hours to prepare a bacterial culture medium. On the next day, 12 test strains were suspended using sterile water according to McFarland standard turbidity of 0.5 to prepare a bacterial culture medium. BMeS-pA was dispensed into a 96 well round bottom microplate after dilution in a 2-fold step in the concentration range of 100 μM to 0.39 μM using CA-MHB. The prepared bacterial culture medium is diluted 1/10 and inoculated by 10 μL into a medium containing BMeS-pA for each concentration (final bacterial concentration 5 x 10 ⁵ CFU/mL). Thereafter, the cells were cultured for 20 hours in an incubator at 37° C., and the MIC was read by visual observation, and the results are shown in FIG. 5 .

As a result, it was confirmed that BMeS-p-A did not inhibit the growth of 12 strains including multidrug-resistant bacteria at any concentration. These results mean that BMeS-p-A has no antibacterial activity against the 12 species tested. However, these results also imply that BMeS-p-A is not toxic to the 12 species tested.

Example 6. Bacterial biofilm fluorescence staining with BMeS-p-A

It was confirmed that BMeS-p-A could perform fluorescence imaging of the biofilm produced by the strain, and this was done through CLSM observation, and the results are shown in FIG. 6 .

As a control, as in Example 2 above, Staphylococcus aureus sensitive and resistant bacteria The main biofilms were simultaneously fluorescently stained and analyzed for comparison.

Specifically, each strain is inoculated with 0.1% strain by putting 2 mL of TSB medium containing 1% glucose on a confocal dish from the strain culture medium cultured overnight in TSB medium. For effective biofilm formation of each strain, it was cultured at 37°C for 48 hours. After that, the supernatant was completely removed, washed three times slowly with phosphate buffered saline, and 100 μM BMeS-p-A as in Example 1 was incubated with shaking at 37° C. for 1 hour. After the reaction, it was thoroughly washed with phosphate buffered saline to remove BMeS-p-A, heat-fixed, and observed by CLSM.

As a result, as shown in FIG. 6, it was confirmed that BMeS-p-A selectively fluorescently stained biofilms formed by Acinetobacter baumani sensitive and resistant strains. On the other hand, fluorescence was not observed in biofilms produced by Staphylococcus aureus susceptible and resistant strains.

Example 7. Bacterial biofilm fluorescence imaging application study by BMeS-p-A

Based on the fact that BMeS-p-A selectively fluorescently stains the biofilm produced by Acinetobacter baumani, it is confirmed whether fluorescence imaging of the biofilm produced by artificially contaminating the urinary catheter where the formation of the bacterial biofilm is a problem is possible. , and the results are shown in Figure 7.

Acinetobacter baumani sensitive and resistant strains capable of selectively fluorescently stained with BMeS-p-A were used, and Staphylococcus aureus sensitive and resistant strains were used as controls.

Specifically, a sterilized ureteral catheter (double pigtail urethral stent) is cut to 2.5 cm and placed in a container for fluorescence measurement containing TSB medium containing 1.0% glucose. Bacteria cultured overnight were inoculated with 0.1% (about 1 x 10 ⁷ CFU/mL) and incubated for 48 hours at 37°C. After incubation, the resulting biofilms were stained in two ways. First, crystal violet was used to check the presence or absence of biofilm formation on the surface of the urinary catheter. Before staining the biofilm on the surface of the urinary catheter, the urinary catheter coated with the biofilm formed by bacteria was carefully washed with phosphate buffered saline and then stained in 1.0% crystal violet solution for 10 minutes. The dye was eluted by gently washing the dyed catheter and shaking it for 20 minutes using 33% glacial acetic acid. The absorbance of the eluted crystal violet was measured at 600 nm using a microplate reader. The second used BMeS-pA for selective biofilm fluorescence imaging. The urinary catheter was stained in the same manner as in Example 6 to perform fluorescence imaging by staining with BMeS-pA. Urinary catheters stained with BMeS-pA were observed using confocal fluorescence microscopy (CLSM).

First, as a result of observing the presence or absence of biofilm formation on the surface of the urinary catheter by crystal violet, it was confirmed that all surfaces of the urinary catheter were stained with crystal violet regardless of the type of strain. Through this, it was found that each strain effectively formed a biofilm and coated the surface of the urinary catheter (FIG. 7a). On the other hand, as a result of observing the fluorescence imaging of the urinary catheter by BMeS-p-A, it was found that the urinary catheter coated with a biofilm formed by Acinetobacter baumanii sensitive and resistant bacteria can selectively perform fluorescence imaging by BMeS-p-A. confirmed (Fig. 7b). On the other hand, fluorescence images could not be observed in the urinary catheter coated with the biofilm formed by the control group Staphylococcus aureus.

The description of the present invention described above is for illustrative purposes, and those skilled in the art can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. There will be. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting.

Claims (12)

A reagent composition for selectively detecting Acinetobacter baumannii containing a compound represented by Formula 2 below:
[Formula 2]

delete The reagent composition according to claim 1, wherein the Acinetobacter baumannii is multi-drug resistant Acinetobacter baumannii . According to claim 3,
A reagent composition, characterized in that the Acinetobacter baumannii is carbapenem-resistant Acinetobacter baumannii . According to claim 1,
Reagent composition, characterized in that for selectively detecting the biofilm (biofilm) produced by the Acinetobacto baumani. According to claim 5,
A reagent composition, characterized in that the biofilm is formed on a medical device. According to claim 6,
A reagent composition, characterized in that the medical device is a urinary catheter. A kit for diagnosing infection of Acinetobacter baumannii , comprising a compound represented by Formula 2 below:
[Formula 2]

Selective detection method of Acinetobacter baumannii , comprising the step of contacting a compound represented by Formula 2 with an analyte:
[Formula 2]

According to claim 9,
The method further comprising the step of observing the fluorescence of the analyte. According to claim 9,
The method of claim 1, wherein the analysis target is a medical device. According to claim 11,
The method of claim 1 , wherein the medical device is a urinary catheter.

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