Composition for detecting antibiotic-resistant bacteria and use thereof

Abstract

The present invention relates to a composition for detection of antibiotic-resistant bacteria and a composition for diagnosis or prognosis of bacterial infectious diseases. The composition for detection according to one aspect has excellent species discrimination on the basis of high target specificity, and thus can be advantageously used to detect antibiotic-resistant bacteria and also to diagnose bacterial infectious diseases caused thereby.

Description

Composition for detecting antibiotic-resistant bacteria and use thereof

[0001] The present invention relates to a composition for detecting antibiotic-resistant bacteria and its use.

[0002] Although antibiotics are considered to have made the greatest contribution to extending human lifespan, their social costs are increasing due to the continuous emergence of resistant bacteria, such as multidrug resistant microbes (MDR) that cannot be controlled by multiple antibiotics.

[0003] Traditional methods for detecting antibiotic-resistant bacteria rely on culture to determine the antibiotic's inhibitory effect. However, culture-based detection methods require 3 to 5 days, making them unsuitable for rapid diagnosis in cases such as bacteremia. To develop a faster and more accurate detection technology for antibiotic-resistant bacteria, we have previously developed a bacterial identification technique using a fluorescent probe signal detection method that does not require culture (Lee, Hyun, Kim, et al., Small Methods, 2022). This method uses a DNA probe specific to the 16S rRNA sequence to identify bacteria. However, the DNA probe must be longer than 20 bp to ensure selectivity, and the charge characteristics of DNA require extended hybridization and washing times, which still present technical limitations.

[0004] The present researchers sought to discover an improved detection composition that overcomes the above limitations, and completed the present invention by confirming the efficacy of an uncharged peptide-bond-based PNA probe.

[0005] One aspect provides a composition for detecting antibiotic-resistant bacteria, comprising a probe comprising any one base sequence selected from the group consisting of sequence numbers 1 to 10 or a complementary base sequence thereof.

[0006] Another aspect provides a kit for detecting antibiotic-resistant bacteria comprising the above-detected composition.

[0007] Another aspect provides a method for detecting antibiotic-resistant bacteria, comprising the step of detecting a mecA gene using the detection composition in a sample isolated from an organism.

[0008] Another aspect provides a composition for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease, comprising the above-detected composition.

[0009] Another aspect provides a kit for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease comprising the above-detected composition.

[0010] Another aspect provides a method for providing information for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease, comprising the step of detecting a mecA gene using the detection composition in a sample isolated from an individual.

[0011] One aspect is to provide a composition for detecting antibiotic-resistant bacteria, comprising a probe comprising any one base sequence selected from the group consisting of sequence numbers 1 to 10 or a complementary base sequence thereof.

[0012] Another aspect is to provide a kit for detecting antibiotic-resistant bacteria comprising the above-detected composition.

[0013] As used herein, the term "antibiotic resistance" refers to the ability of microorganisms to survive exposure to antibiotics. This is a survival strategy for bacteria to survive the attack of antibiotics. When antibiotics are administered, bacteria susceptible to the antibiotic die, while some resistant bacteria, or "antibiotic-resistant bacteria," survive and proliferate under selective pressure. Therefore, antibiotic use leads to resistance, and some resistance genes can be horizontally transferred, spreading resistance to other bacteria and causing significant problems for the host.

[0014] Antibiotic resistance mechanisms largely involve methods that prevent the antibiotic from reaching its target in sufficient concentration or alter or bypass its target. Examples of resistance mechanisms include drug inactivation, alteration of the target site, decreased uptake, immediate efflux pump removal, and alteration of the metabolic pathway.

[0015] The antibiotic-resistant bacteria to be detected in the present invention may be Staphylococcus aureus.

[0016] Staphylococcus aureus can be divided into methicillin-resistant Staphylococcus aureus (MRSA) or methicillin-susceptible Staphylococcus aureus (MSSA) depending on the presence or absence of the mecA gene, and the antibiotic-resistant bacteria to be detected in the present invention may be, more specifically, MRSA. MRSA is Staphylococcus aureus that is resistant to methicillin and other β-lactam antibiotics, and refers to all Staphylococcus aureus that is resistant not only to methicillin but also to penicillinase-resistant penicillins (PRPs) such as oxacillin, nafcillin, and cloxacillin.

[0017] The most fundamental mechanism by which MRSA develops resistance to penicillin-like receptors (PRPs), such as methicillin, is a decrease in the affinity of the drug's target. Specifically, MRSA can continue to proliferate unaffected by beta-lactam antibiotics by producing PBP2a (penicillin-binding protein 2a), a receptor with very low binding affinity for beta-lactam antibiotics, including PRP. PBP2a (also called PBP2') is expressed by the mecA gene present on the MRSA chromosome.

[0018] As used herein, the term “detection” means confirming the presence and / or expression level of mRNA of genes in a biological sample.

[0019] The term "probe" as used herein refers to a substance capable of specifically binding to a target substance to be detected within a sample, and through said binding, the presence of the target substance within the sample can be specifically confirmed. The probe has a base sequence that is partially or completely complementary to the target nucleic acid base sequence.

[0020] The type of probe molecule is not limited to a material commonly used in the art, but may preferably be PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA or DNA, and may be a concept that includes all biomaterials derived from or similar to living organisms or manufactured in vitro.

[0021] The detection composition of the present invention may preferably be a PNA probe. As used herein, the term "peptide nucleic acid (PNA)" refers to a DNA-like substance in which nucleic acid bases are linked by peptide bonds rather than phosphate bonds, and whose basic backbone is composed of polyamide. It is one of the artificially synthesized genetic recognition substances, such as LNA (Locked nucleic acid) or MNA (Mopholino nucleic acid), and was first synthesized by Nielsen et al. in 1991.

[0022] PNA possesses excellent affinity and selectivity for target genes. Its neutral structure, free of the negative charge characteristic of the DNA backbone, allows for strong binding. Because DNA probes are electrically negatively charged, there is a repulsive force between like-pole DNA molecules when they bind to form a double helix structure. However, PNA probes possess a neutral, uncharged backbone, so there is no electrical repulsion when binding to DNA or RNA. PNA probes can exhibit excellent species specificity.

[0023] PNA probes can penetrate microorganisms more quickly than conventional DNA probes, recognize ribosomal RNA sequences, and significantly shorten the time to detect or identify specific microbial species. According to one specific example, when a PNA probe is used, the time to detect or identify a specific microbial species can be significantly reduced compared to when a conventional DNA probe is used. Specifically, when a DNA probe is used, low detection efficiency is exhibited even after hybridization for about 12 hours, whereas when a PNA probe is used, significantly increased detection efficiency can be exhibited with only hybridization for about 1 to 10 minutes. The above-mentioned contents are described in detail in Korean Patent Publication No. 10-2533260, the contents of which may be incorporated herein by reference in their entirety.

[0024] In addition, PNA, which has a peptide backbone, is closer to a protein (peptide) than a nucleic acid chemically, so it has the advantage of being highly stable against nucleases and not being degraded by existing restriction enzymes. In addition, it has high thermal and chemical properties and stability, such as a melting temperature that is about twice as high as DNA, so it is easy to store and has the advantage of not being easily degraded. PNA forms a double strand through a hybridization reaction with a natural nucleic acid of complementary base sequence, and when the length is the same, a PNA / DNA duplex is more stable than a DNA / DNA duplex, and a PNA / RNA duplex is more stable than a DNA / RNA duplex. Since the degree to which the double strand of PNA becomes instable due to a single base mismatch is high, the ability to detect single nucleotide polymorphisms (SNPs) is superior to that of natural nucleic acids. Furthermore, because it is artificially synthesized, it offers the advantage of being easily modified for secondary applications, allowing for the easy use of detection markers or useful molecules. By linking specific molecules or functional groups to the chemical structure of PNA, its physicochemical properties can be modified to suit specific applications.

[0025] The above detection composition may be intended to detect the mecA gene.

[0026] The above detection composition may comprise a probe comprising a sequence capable of hybridizing with a portion of the mecA gene.

[0027] The above detection composition may include a probe comprising a sequence capable of hybridizing with 10 to 20 consecutive base sequence regions among the base sequences of the mecA gene.

[0028] The above detection composition may be composed of 10 to 20 consecutive base sequences among the complementary base sequences of the mecA gene of SEQ ID NO: 11, and may include a probe including any one of the base sequences of SEQ ID NOs: 1 to 10.

[0029] The above detection composition may include a probe including one or more base sequences selected from the group consisting of the 188th to 202nd, 235th to 249th, 318th to 332nd, 513th to 527th, 920th to 934th, 1352nd to 1366th, 1412th to 1426th, 1547th to 1561st, 1603rd to 1617th, and 1789th to 1803rd nucleotide sequences from one end of the mecA gene of SEQ ID NO: 11, or a complementary base sequence thereof.

[0030] Specifically, the detection composition may include a probe comprising a base sequence from the 318th to the 332nd base sequence at one end of the mecA gene of SEQ ID NO: 11 or a complementary base sequence thereof.

[0031] Specifically, the detection composition may include a probe comprising a base sequence from the 920th to the 934th base sequence at one end of the mecA gene of SEQ ID NO: 11 or a complementary base sequence thereof.

[0032] Specifically, the detection composition may include a probe comprising a base sequence from the 1412th to the 1426th base sequence at one end of the mecA gene of SEQ ID NO: 11 or a complementary base sequence thereof.

[0033] The above detection composition may include a polynucleotide comprising a nucleotide sequence having at least 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% homology to any one of the base sequences of SEQ ID NOs: 1 to 10 or a complementary base sequence thereof. It is obvious that a polynucleotide sequence having a biological activity substantially identical to or corresponding to the sequence having homology thereto is also included in the scope of the present application, even if it has a polynucleotide sequence in which a part of the sequence is deleted, modified, substituted, or added.

[0034] Specifically, the detection composition may include a polynucleotide comprising a nucleotide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% homology to the base sequence of the sequence number 3 or a complementary base sequence thereof.

[0035] Specifically, the detection composition may include a polynucleotide comprising a nucleotide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% homology to the base sequence of the sequence number 5 or a complementary base sequence thereof.

[0036] Specifically, the detection composition may include a polynucleotide comprising a nucleotide sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% homology to the base sequence of the sequence number 7 or a complementary base sequence thereof.

[0037] In the present invention, the term "hybridization" refers to the formation of a double-stranded nucleic acid by hydrogen bonding between single-stranded nucleic acids with complementary base sequences, and is used in a similar sense to annealing. However, in a broader sense, it may include cases where the base sequences between two single-stranded nucleic acids are completely complementary (perfect match), as well as exceptional cases where some base sequences are not complementary (mismatch).

[0038] The analysis method of the hybridization reaction can be performed by a method known in the art, such as histochemistry, immunohistochemistry, immunofluorescence, chromogenic in situ hybridization, fluorescent in situ hybridization (FISH), etc. Specifically, in the detection step, the presence or absence of antibiotic-resistant bacteria or the type of antibiotic-resistant bacteria can be determined by comparing the signal of the portion where the signal emitted from the probe hybridized with the antibiotic-resistant bacteria exists with the signal of the other portion. In addition, the concentration of antibiotic-resistant bacteria contained in the sample can be obtained by comparing with a standard sample containing antibiotic-resistant bacteria at a known concentration in advance.

[0039] The term "FISH (Fluorescent in situ Hybridization) analysis method" used in this specification refers to an analysis method that detects a specific DNA sequence in a chromosome using a probe attached to a fluorescent molecule. It is used for the purpose of identifying the presence or absence of a specific base sequence, and the term "cross-linking," "hybridization," or "cross-linking" refers to an inspection process that binds a probe to a chromosome, and the probe bound to the chromosome fluoresces when exposed to ultraviolet rays, indicating the presence or absence and location of the corresponding sequence. After binding a label to the DNA probe, the DNA of the cell to be tested is denatured to form a single strand and then reacted. At this time, the DNA probe reacts with the target, and after washing, when a fluorescent substance capable of binding to the label is added, the DNA probe exhibits fluorescence at the binding site.

[0040] The above probes can be chemically synthesized using the phosphoramidite solid support method or other well-known methods. These nucleic acid sequences can also be modified using many means known in the art. Non-limiting examples of such modifications include methylation, capping, substitution with one or more homologs of a natural nucleotide, and modification between nucleotides, such as modification with uncharged linkers (e.g., methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.) or charged linkers (e.g., phosphorothioate, phosphorodithioate, etc.).

[0041] The above probe may be labeled for easier detection. That is, it may be characterized by having a reporter or a quencher linked to both ends. When the distance between the reporter and the quencher is close, the signal generation of the probe is suppressed, and as the distance between the reporter and the quencher increases, the signal intensity increases. Generally, when the probe hybridizes with a complementary base sequence, the distance between the reporter and the quencher is the farthest, so a specific base sequence can be detected through signal generation or increased signal intensity. By linking the probe to the detection label (reporter, quencher, etc.), the microorganism can be visualized (or imaged), and the microorganism can be detected or identified. By using a probe linked to the detection label, the hybridization of the probe and the microorganism, and the dissociation of the probe hybridized with the microorganism and the detection label can be easily confirmed by simple imaging due to the change in the signal of each detection label.

[0042] The reporter may be characterized by being at least one fluorescent substance selected from the group consisting of fluorescein, fluorescein chlorotriazinyl, rhodamine green, rhodamine red, tetramethylrhodamine, FITC, Oregon green, Alexa Fluor, FAM, JOE, ROX, HEX, Texas Red, TET, TRITC, TAMRA, cyanine series dyes, and thiadicarbocyanine dyes.

[0043] The above quencher may be characterized by being at least one selected from the group consisting of Dabcyl, TAMRA, Eclipse, DDQ, QSY, Blackberry Quencher, Black Hole Quencher, Qxl, Iowa black FQ, Iowa black RQ, and IRDye QC-1.

[0044] In the present invention, the “kit” may optionally include reagents necessary for performing a target nucleic acid amplification reaction (e.g., polymerase chain reaction), such as a buffer, DNA polymerase, a DNA polymerase cofactor, and deoxyribonucleotide-5-triphosphate (dNTP). Optionally, the kit of the present invention may also include various oligonucleotide molecules, reverse transcriptase, various buffers and reagents, and antibodies that inhibit DNA polymerase activity. Furthermore, the optimal amount of reagents to be used in a particular reaction of the kit can be readily determined by one of ordinary skill in the art having learned the disclosure herein. Typically, the device of the present invention may be manufactured as a separate package or compartment containing the aforementioned components.

[0045] In one embodiment, the kit may comprise a compartmented carrier means for containing a sample, a container containing a reagent, and a container containing a PNA probe.

[0046] The carrier means is suitable for containing one or more containers, such as bottles or tubes, each containing independent components used in the method of the present invention. Given the present disclosure, one skilled in the art can readily dispense the required formulations within the containers.

[0047] In the present invention, "sample" includes various samples, and preferably, a biological sample is analyzed using the method of the present invention. The sample may include a biological sample, an environmental sample, or a food sample. The biological sample may refer to a sample obtained from a biological subject, including a sample of biological tissue or body fluid obtained in vivo or in vitro. Specifically, the biological sample may be, but is not limited to, a body fluid (e.g., blood, plasma, serum, saliva, sputum, or urine), organ, tissue, fraction, or cell isolated from a mammal, including a human. In addition, the biological sample may include an extract from the biological sample, for example, antibodies, proteins, etc. from a biological fluid (e.g., blood or urine). The environmental sample may include a water sample or a soil sample. The sample may contain microorganisms, i.e., antibiotic-resistant bacteria.

[0048] Another aspect provides a method for detecting antibiotic-resistant bacteria, comprising the step of detecting the mecA gene in a sample isolated from an organism using the detection composition. The description of the composition and kit also applies to the detection method.

[0049] The above detection method may additionally include a step of contacting the PNA probe with antibiotic-resistant bacteria in the sample to hybridize with the antibiotic-resistant bacteria.

[0050] The above detection method may additionally include a step of detecting a signal emitted from a PNA probe hybridized with an antibiotic-resistant bacteria in the sample.

[0051]

[0052] Another aspect is to provide a composition for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease comprising the above-detected composition.

[0053] Another aspect is to provide a kit for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease comprising the above-detected composition.

[0054] Another aspect provides a method for providing information for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease, comprising the step of detecting a mecA gene using the detection composition in a sample isolated from an individual.

[0055] The description of the above detection composition, kit and / or method also applies to the above diagnosis or prognosis prediction composition, kit and / or method.

[0056] In this specification, “antibiotic-resistant bacterial infectious disease” may refer to any type of disease caused by an antibiotic-resistant bacterial infection. The antibiotic-resistant bacterial infectious disease may be bacteremia, sepsis, septic shock, pneumonia, food poisoning, bacterial dysentery, impetigo, purulent disease, abscess and ulcer, boil, pyoderma, folliculitis, dermatitis, wound infection, endocarditis, osteomyelitis, and / or enteritis.

[0057] The above antibiotic-resistant bacterial infectious disease may be a disease caused by methicillin-resistant Staphylococcus aureus (MRSA), i.e., MRSA infection.

[0058] As used herein, the term "diagnosis" refers to confirming the presence or characteristics of a pathological condition. For the purposes of the present invention, diagnosis may refer to determining whether a bacterial infectious disease has developed.

[0059] The term "prognosis" in this specification refers to a prediction regarding the progression or recovery of a disease, and refers to a prospect or preliminary assessment. For the purpose of the present invention, prognosis means determining whether treatment success, survival, recurrence, metastasis, drug responsiveness, resistance, etc. of a subject after treatment of a bacterial infectious disease are present. In other words, it refers to an expectation regarding the medical outcome (e.g., long-term survival possibility, disease-free survival rate, etc.), and includes a positive prognosis (positive prognosis) or a negative prognosis (negative prognosis). The negative prognosis includes disease progression or mortality such as recurrence, metastasis, drug resistance, etc., and the positive prognosis includes disease remission such as the absence of disease, and disease improvement or stabilization such as tumor regression.

[0060] The composition for detection according to the aspect has high target specificity and homogeneous labeling, and thus has high species discrimination ability for individual bacteria, so it can be usefully used not only for detecting antibiotic-resistant bacteria, but also for diagnosing bacterial infectious diseases caused by antibiotic-resistant bacteria.

[0061] Figure 1 is a schematic diagram showing the design positions of PNA probes of sequence numbers 1 to 10 in order.

[0062] Figure 2 is an image comparing and analyzing the species discrimination ability of PNA probes for individual bacteria according to one specific example.

[0063] Figure 3 is a graph comparing and analyzing the species discrimination ability of PNA probes for individual bacteria according to one specific example.

[0064] Figure 4 is a graph showing the results of MSSA tests and MRSA tests of a PNA probe according to one specific example.

[0065] The following examples are provided for more detailed description. However, these examples are provided solely to illustrate one or more specific examples, and the scope of the present invention is not limited to these examples.

[0066]

[0067] Example 1. Preparation of PNA probe

[0068] According to one specific embodiment of the present invention, an antibiotic resistance mRNA imaging probe was designed to select only conserved regions unaffected by known mutations in the corresponding gene, and to have target specificity when mismatches of 2 bp or more with the representative bacterial genome (genomic DNA) were allowed. The design positions of the 10 finally selected detection PNA probes are sequentially shown in Fig. 1, and specifically, the probes were designed as shown in Table 1 below.

[0069]

[0070] Number sequence mecA-01 (SEQ ID NO: 1) CTGAACGTCCGATAAmecA-02 (SEQ ID NO: 2) AACATTCAGGATCGTmecA-03 (SEQ ID NO: 3) TAACATTGATCGCAAmecA-04 (SEQ ID NO: 4) AGGCATCGTTCCAAAmecA-05 (SEQ ID NO: 5) ATACAATCGCACATAmecA-06 (SEQ ID NO: 6) GTAATATCGACTTAAmecA-07 (SEQ ID NO: 7) CACTCGAATTAGGCAmecA-08 (SEQ ID NO: 8) ATTCAGGTTACGGACmecA-09 (SEQ ID NO: 9) TATAGCGCATTAGAAmecA-10 (SEQ ID NO: 10) AAATCCGGTACTGCA

[0071]

[0072] Experimental Example 1. Evaluation of the species discrimination ability of PNA probes

[0073] To confirm that the PNA probe according to one specific example has high species discrimination power, the following Fluorescence in situ hybridization (FISH) experiment was performed.

[0074] Specifically, experiments were conducted on seven bacterial species, including Klebsiella pnuemoniae, Proteus mirabilis, Bacillus subtilis, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. All bacterial species were cultured on LB agar plates at 37°C for 20 h, and then a single colony was inoculated into 3 ml of LB medium and grown at 37°C. The culture was stopped when the absorbance at 600 nm reached 0.4–0.6 (log phase) using a UV-Vis spectrophotometer every 30 min. The cultured bacterial species were concentrated 2–4 times, and 10 μL was placed on a poly-L-lysine-coated confocal microscope slide. Slides for confocal microscopy were prepared by sonicating with 1 M nitric acid at room temperature for 1 hour, washing three times with deionized water, drying with nitrogen, soaking the slides in a 0.01% poly-L-lysine solution at 37°C for 1 hour, washing three times with deionized water, and completely drying in a vacuum chamber for 1 hour.

[0075] After incubating the slides at room temperature for 30 minutes, they were first washed with nuclease-free water, then bacterial cells were fixed with 200 μL of a 3.7% formaldehyde solution at room temperature for 30 minutes, and washed three times with nuclease-free water. Bacterial cells were permeabilized with 200 μL of a 5 mg / ml lysozyme solution dissolved in nuclease-free water for 30 minutes at room temperature. Afterwards, the PNA probe was hybridized with 100 μL solution containing 10% w / v dextran sulfate, 10 mM NaCl, 30% formamide, 0.1% w / v sodium pyrophosphate, 0.1% Triton X-100, 50 mM Tris-HCl (pH 7.5), 0.2% w / v polyvinylpyrrolidone, 0.2% w / v Ficoll, 5 mM disodium EDTA, and 200 nM PNA probe of Example 1 in a humidified chamber at 55°C for 30 min. The sample was washed twice for 15 min at 55°C in 200 μL wash solution containing 15% formamide, 5 mM Tris-base (pH 10), 15 mM NaCl, and 0.1% Hoechst 33342.

[0076] Images were acquired using a Zeiss LSM 880 confocal microscope with a 40x water immersion objective (Carl Zeiss, NA 1.0). 561 nm, 633 nm, and 405 nm lasers were used for the Cy3, Cy5, and DAPI channels, respectively. Cell boundaries were determined from the DAPI channel images, and the fluorescence signal of each cell was measured as the average of the background levels within the detected boundaries. The intercellular distribution of this average was plotted as a scatter plot and histogram, and statistical values ​​such as the mean, variance, and confidence interval were extracted, and crosstalk between species was calculated, and the results are shown in Figs. 2 and 3.

[0077] As shown in FIGS. 2 and 3, the PNA probe according to one specific example was observed to have high target specificity for S. aureus, but hardly bind to other bacterial species with small sequence variations in 16S rRNA.

[0078] These results indicate that the PNA probe has high target specificity for Staphylococcus aureus and high species discrimination power for individual bacteria.

[0079]

[0080] Experimental Example 2. Evaluation of the ability of PNA probes to detect mecA gene expression signals.

[0081] To evaluate the detection ability of the mecA gene expression signal of the PNA probe according to one specific example, the MSSA test and the MRSA test were performed.

[0082] Specifically, the culture of methicillin-resistant Staphylococcus aureus (MRSA), the culture of methicillin-susceptible Staphylococcus aureus (MSSA), and the preparation of slides for imaging were carried out in the same manner as in Experimental Example 1, except that methicillin was added at a concentration of 64 μg / mL before bacterial attachment and incubated for 1 hour. The results of the MSSA test and MRSA test are shown in Fig. 4.

[0083] As shown in Fig. 4, it was confirmed that the PNA probe according to one specific example has excellent MRSA detection ability.

[0084] The results above indicate that the PNA probe according to one specific example has excellent detection capability of the expression signal of the mecA gene, which is an antibiotic resistance gene, and can be used for the detection of antibiotic-resistant bacteria and the diagnosis of related diseases.

[0085]

[0086] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. A composition for detecting antibiotic-resistant bacteria, comprising a probe comprising any one base sequence selected from the group consisting of sequence numbers 1 to 10 or a complementary base sequence thereof.

2. A composition for detection according to claim 1, wherein the probe is a PNA probe.

3. A composition for detection according to claim 1, wherein the antibiotic-resistant bacteria is methicillin-resistant Staphylococcus aureus (MRSA).

4. A kit for detecting antibiotic-resistant bacteria comprising a composition according to any one of claims 1 to 3.

5. A method for detecting antibiotic-resistant bacteria, comprising the step of detecting a mecA gene using a composition according to any one of claims 1 to 3 in a sample isolated from an organism.

6. A composition for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease, comprising the composition of any one of claims 1 to 3.

7. A kit for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease, comprising a composition according to any one of claims 1 to 3.

8. A method for providing information for diagnosing or predicting the prognosis of an antibiotic-resistant bacterial infectious disease, comprising the step of detecting a mecA gene using a composition according to any one of claims 1 to 3 in a sample isolated from an organism.

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