KR102318149B1 - Biomarker composition for discrimination of meticillin resistant or sensitive Staphylococcus aureus clone type - Google Patents

Abstract

The present invention relates to a biomarker composition for discriminating methicillin-resistant or sensitive Staphylococcus aureus clonal type and its use, and more particularly, for the genotype of a hospital-borne infection (ST5) and a genotype of a regionally-borne infection (ST72) among Staphylococcus aureus. Provides a biomarker composition that is differentially expressed in a total of four types of MSSA (Methicillin-sensitive SA) and MRSA (Methicillin-resistant SA), respectively, and the biomarker composition of the present invention is a gene group showing differences in expression according to the clone type. Through the combination, the diagnosis of methicillin-resistant or sensitive Staphylococcus aureus infection and the clone type can be confirmed, so it can be used for clinically simple and effective identification methods and kits for identification.

Description

Biomarker composition for discrimination of methicillin resistant or sensitive Staphylococcus aureus clone type

The present invention relates to a biomarker composition for discriminating methicillin-resistant or sensitive Staphylococcus aureus clonetypes and uses thereof.

Staphylococcus aureus is a major pathogen that can cause both mild and severe infections in various parts of the body. Among them, methicillin-resistant Staphylococcus aureus (MRSA) causes nosocomial infections It is well known as one of the most common causative agents.

MSSA (methicillin-sensitive Staphylococcus aureus) refers to Staphylococcus aureus showing sensitivity to penicillinase-resistant penicillin (PRP) in a relative meaning to MRSA. The criterion to distinguish MRSA from MSSA is the presence or absence of SCCmec containing mecA. Due to these genotype differences, MRSA shows resistance to PRP, and MSSA shows sensitivity to phenotype differences. However, other than the simple difference that MRSA is resistant to PRP and MSSA is not resistant to PRP, there has been no report on any other differences between the two strains in the phenotype related to PRP resistance. , the prognosis may vary. Therefore, accurate and rapid bacterial identification and antimicrobial susceptibility test are essential for proper selection of antimicrobial agents and management of nosocomial infections.

MRSA is an important pathogen causing nosocomial infections (healthcare-associated (HA)-MRSA), but MRSA infections are also common in community-acquired (CA)-MRSA. According to previous reports, the frequency of CA-MRSA in Korea was found to be 112 (5.9%) out of a total of 1,900 MRSA patients. The patients were mainly skin infections or soft tissue infections, and 64% showed multidrug resistance. Accordingly, as a result of molecular epidemiologic analysis of CA-MRSA strains, staphylococcal chromosomal cassette mec (SCCmec) type IVa was the most common SCCmec type, and the sequence type was ST72 the most. However, as for HA-MRSA, according to previous reports, as a result of analyzing the MLST results of MRSA isolated from intensive care unit patients at a university hospital in 1996 and 2004, ST5 prevailed in 1996, but ST239 was the most common type in 2004. has been reported to have changed to

Conventional methods for diagnosing MRSA include: 1) culturing the patient's blood for more than 24 hours and isolating the bacteria to perform an antimicrobial sensitivity test, or 2) culturing the bacteria and directly in the colony to MRSA with a specific gene (mec-A gene). ) or 3) MRSA-specific protein (PBP-2a) is detected. However, there are limitations to the MRSA diagnosis method because it takes more than 24 hours for blood culture, there are cases where bacterial culture is not possible, and there are cases where colonies are not formed.

Republic of Korea Patent Registration No. 10-1779038 (Registered on September 11, 2017)

In order to solve the above problems, the present invention is to provide a biomarker composition for methicillin-resistant or sensitive Staphylococcus aureus clone type discrimination.

The present invention provides aacA (Aminoglycoside phosphotransferase APH(3'))(Accession: D3X7Z4), ermA (23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(B))(Accession: Q7DIC2), ant(9) )(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), NCTC11940_00344 (Protein of uncharacterized function (DUF3644))(Accession: A0A2X3P763), CSC83_13340 (Capsular biosynthesis protein)(Accession: A0A2840_00987) (Accession: A0A2S) Accession: A0A167QYC2), hmp (Flavohemoprotein) (Accession: A0A1D4MV18), SAP027A_009 (Antiseptic resistance protein) (Accession: D2J776), BN1321_390017 (Uncharacterized protein) (Accession: A0A0U1MRY5), , mecR1 (Methicillin resistance regulatory sensor-transducer MecR1) (Accession: A0A2Y2RBM2), cydA (Cytochrome d ubiquinol oxidase subunit I) (Accession: A0A167UFF7), lytS (Autolysis histidine kinase LytS) (01017PHage9), related protein) (Accession: A0A0U1MY60), SAMEA170867 4_03678 (Cell division inhibitor)(Accession: A0A2W2UEW9), capC (Tyrosine-protein phosphatase)(Accession: A0A0U1MX93), BTN44_14990 (Lipoprotein)(Accession: A0A0D3QB23), C0J46_00K500 (Ribok: rbs0A)(Accession: Abs0A)(Accession: Abs0A) (Accession: A0A2Y2LM71), ERS140147_00739 (Membrane protein) (Accession: A0A077VLJ7), bglA (6-phospho-beta-glucosidase) (Accession: A0A0D6GG52), pyrR (Bifunctional protein PyrR) (Accession: W8U6N8), vraR (DNA-binding), vraR (DNA-binding) response regulator)(Accession: D9IFM7), yxeP_2 (Carboxypeptidase, M20(D) family)(Accession: A0A0U1MEH9), orfX (Ribosomal RNA large subunit methyltransferase H)(Accession: A0A068W8S1), aroA (3-phosphoshikimate)(1-carboxyvinyltransferase)( Accession: A0A133PU31), queF (NADPH-dependent 7-cyano-7-deazaguanine reductase) (Accession: A0A077V3Q2), ydfH (Gluconate operon transcriptional repressor) (Accession: A0A1D4Q3C3), C7Q89_10110 (Abortive phage resistance protein) (Abortive phage resistance protein) , rpsC (30S ribosomal protein S3) (Accession: A0A1Q8DBU3), traP (Signal transduction protein TRAP)(Accession: A0A0U1N0V2), BSZ10_08125 (Alkaline-shock protein)(Accession: A0A1Q8DDB2), SAP059A_003 (Antiadhesin Pls, binding to squamous nasal epithelial cells)(Accession: D2) 2-oxobutanoate hydroxymethyltransferase) (Accession: A0A0Z1DQN8), rpsO (30S ribosomal protein S15) (Accession: A0A077VJU6), SAMEA1708674_01891 (Enterotoxin) (Accession: A0A1D4MQV2), ndk (Nucleoside) chain alcohol dehydrogenase)(Accession: A0A1D4IQF8), sbi (Immunoglobulin-binding protein sbi)(Accession: A0A0U1MSJ9), parB_2 (Chromosome partitioning protein ParB)(Accession: A0A0D3Q4U5), CSC83_07060 (Uncharacterized protein)_2 (Uncharacterized protein)_2) Ribosomal-protein-L7p-serine acetyltransferase)(Accession: A0A1D4MIM0), BTN44_15570 (Putative transposase)(Accession: B6V383), BN1321_380004 (DUF961 domain-containing protein)(Accession: A0A0U1MRA0), C7Q89_00130 (Protein-disulfide isomerase) (Protein-disulfide isomerase) ccession: A0A2W2RV36), truB_2 (Fibronectin-binding protein A)(Accession: A0A0U1MF71), ecsA_2 (Lantibiotic transport ATP-binding protein)(Accession: A0A0U1MEQ0), SAMEA1708674_03752 (Cell division protein (Putative))(Accession: A0A0U1MEQ0)(Cell division protein (Putative)) (Chaperone protein ClpB)(Accession: A0A2W3CJ21), argG (Argininosuccinate synthase)(Accession: A0A0D6WAS8), saeR (Response regulator SaeR)(Accession: A0A0U1MFP7), EP54_11640 (DegV domain-containing protein) 6HNG6HNG0A0 (Accession: A0A0D6WAS8) DNA-binding response regulator)(Accession: A0A2Y9TRS2), HMPREF3211_00240 (GntR family transcriptional regulator)(Accession: A0A133QB66), BSZ10_11980 (Transposon DNA-invertase)(Accession: A0A1Q8DBUN86), OX=1280 (Uncharacterized protein)(Accession: A0A2S1) , mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) and mnhE (Na (+) H (+) antiporter subunit E) (Accession: A0A077VW30) any one or more genes selected from the group consisting of or a protein encoded by the gene It provides a biomarker composition for methicillin-resistant or sensitive Staphylococcus aureus clone type discrimination.

The present invention relates to a probe that specifically binds to any one or more genes in the biomarker composition, a primer for amplifying the gene, an antibody that specifically binds to a protein encoded by the gene, or a binding domain specific to the protein. It provides a kit for discriminating methicillin-resistant or sensitive Staphylococcus aureus clones comprising a peptide having,

In addition, the present invention provides aacA (Aminoglycoside phosphotransferase APH(3'))(Accession: D3X7Z4), ermA (23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(B)) (Accession: Q7DIC2), ant(9)(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), NCTC11940_00344 (Protein of uncharacterized function (DUF3644))(Accession: A0A2X3P763), CSC83_13340 (Capsular biosynthesis protein)(Capsular biosynthesis protein) ), ERS072840_00987 (Phage repressor protein)(Accession: A0A167QYC2), hmp (Flavohemoprotein)(Accession: A0A1D4MV18), SAP027A_009 (Antiseptic resistance protein)(Accession: D2J776), BN1321_U1MRY5) (Uncharacterized protein)(0695: ASC0A)(0695) major capsid protein) (Accession: A0A2S6DEN9), mecR1 (Methicillin resistance regulatory sensor-transducer MecR1) (Accession: A0A2Y2RBM2), cydA (Cytochrome d ubiquinol oxidase subunit I) (Accession: A0A167UFF7), his lytidine kinase (Accession: A0A167UFF7), his lytidine kinase : A0A131JSM9), SAMEA2445518_01017 (Hypothetical phage-related protein) (Accession: A0A0U1MY60), SAMEA1708674_03678 (Cell division inhibitor)(Accession: A0A2W2UEW9), capC (Tyrosine-protein phosphatase)(Accession: A0A0U1MX93), BTN44_14990 (Lipoprotein) (Accession: A0A0D3QB23FHQ), HQ4r (Ribokinase) (Accession: A0A2Y2LM71), ERS140147_00739 (Membrane protein) (Accession: A0A077VLJ7), bglA (6-phospho-beta-glucosidase) (Accession: A0A0D6GG52), pyrR (Bifunctional protein PyrR) (Accession: W8U) DNA-binding response regulator) (Accession: D9IFM7), yxeP_2 (Carboxypeptidase, M20(D) family) (Accession: A0A0U1MEH9), orfX (Ribosomal RNA large subunit methyltransferase H) (Accession: A0A068W8S1), aroshikimate 1-phosphoshikimate carboxyvinyltransferase) (Accession: A0A133PU31), queF (NADPH-dependent 7-cyano-7-deazaguanine reductase) (Accession: A0A077V3Q2), ydfH (Gluconate operon transcriptional repressor) (Accession: A0A1DQ3C3), C7Q89 (AbortiveAccess4Q3C3), C7Q89 : B2Y834), rpsC (30S ribosomal protein S3) (Accession: A0A1Q8DBU3), traP (Signal transduction protein TRAP)(Accession: A0A0U1N0V2), BSZ10_08125 (Alkaline-shock protein)(Accession: A0A1Q8DDB2), SAP059A_003 (AntiadhesinJ Accession2), nasal epithelial cells: panB (3-methyl-2-oxobutanoate hydroxymethyltransferase)(Accession: A0A0Z1DQN8), rpsO (30S ribosomal protein S15)(Accession: A0A077VJU6), SAMEA1708674_01891 (Enterotoxin)(Accession: A0A1D4MQV2), phosphate (Nucle kinase)(Accession: A0A1D4MQV2) ), SAMEA1708674_00628 (Short-chain alcohol dehydrogenase)(Accession: A0A1D4IQF8), sbi (Immunoglobulin-binding protein sbi)(Accession: A0A0U1MSJ9), parB_2 (Chromosome partitioning protein ParB)(Accession: A0A07060) (Uncharacterized protein) (Uncharacterized protein) : A0A2L2EVY2), ydaF_2 (Ribosomal-protein-L7p-serine acetyltransferase)(Accession: A0A1D4MIM0), BTN44_15570 (Putative transposase)(Accession: B6V383), BN1321_380004 (DUF961), (CProte0A0Accession: A0A1D4MIM0) in-disulfide isomerase)(Accession: A0A2W2RV36), truB_2 (Fibronectin-binding protein A)(Accession: A0A0U1MF71), ecsA_2 (Lantibiotic transport ATP-binding protein)(Accession: A0A0U1MEQ0), SAMEA1708674_03752 (Cell division protein (Putative)) Accession: A0A1D3YL80), clpB (Chaperone protein ClpB)(Accession: A0A2W3CJ21), argG (Argininosuccinate synthase)(Accession: A0A0D6WAS8), saeR (Response regulator SaeR)(Accession: A0A0U1MFP7) (Accession: A0A0U1MFP7) (Accession domain-containing protein) : A0A0D6HNG8), RK64_10785 (DNA-binding response regulator)(Accession: A0A2Y9TRS2), HMPREF3211_00240 (GntR family transcriptional regulator)(Accession: A0A133QB66), BSZ10_11980 (Transposon DNA-invertase)(Uncharacterized protein)(Accession: A0A1Q8) ) (Accession: A0A2S1FUN9), mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) and mnhE (Na(+) H(+) antiporter subunit E) (Accession: A0A077VW30) mRNA expression of any one or more genes selected from the group consisting of Checking the level or the expression level of the protein encoded by the gene; and

It provides a method for determining the methicillin-resistant or sensitive Staphylococcus aureus clone type comprising the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control.

The present invention is a biomarker differentially expressed in a total of four types of MSSA (Methicillin-sensitive SA) and MRSA (Methicillin-resistant SA), respectively, for the hospital-borne infection genotype (ST5) and the regionally-borne infection genotype (ST72) among Staphylococcus aureus. With respect to the composition, the biomarker composition of the present invention is clinically convenient because it is possible to diagnose methicillin-resistant or sensitive Staphylococcus aureus infection and confirm the clone type through the combination of the gene group showing the expression difference according to the clone type. It can be used for the production of effective discrimination methods and kits for discrimination.

1 is a view showing the biomarker screening method of the present invention.
Figure 2 shows the change in protein expression pattern according to each sample in the data obtained through mass spectrometry, the protein is quantified through the label-free quantitation method, and the group of proteins is classified into 9 clusters according to the pattern change between the samples.
FIG. 3 is a result of confirming nine clusters of proteins showing the same quantitative expression pattern in eight data sets in which four groups of ST5 MRSA, ST5 MSSA, ST72 MRSA and ST72 MSSA were analyzed twice.
4 is a result showing the accession number, name, gene name, standardized Z score, and log value of the p value of the ANOVA test of 20 proteins with a specific high expression level in ST5 MRSA.
5 is a result showing the accession number, name, gene name, standardized Z score, and log value of the p value of the ANOVA test of 10 proteins with a specifically high expression level in ST72 MRSA.
6 is a result showing the accession number, name, gene name, standardized Z score and log value of the p value of the ANOVA test of 18 proteins with a specific high expression level in ST5 MSSA.
7 is a result showing the accession number, name, gene name, standardized Z score and log value of the p value of the ANOVA test of 10 proteins with a specific high expression level in ST72 MSSA.

Hereinafter, the present invention will be described in more detail.

The present invention provides a total of 4 groups (ST5 MSSA, ST5 MRSA, ST72 MSSA, ST72) for each of the hospital-borne infection genotype (ST5) and the regionally transmitted infection genotype (ST72). Staphylococcus aureus was isolated from the blood of 40 patients (10 in each group) infected with MRSA), and proteins differentially expressed in the 4 groups were discovered using high-resolution mass spectrometry and statistical analysis, thereby completing the present invention.

The present invention provides aacA (Aminoglycoside phosphotransferase APH(3'))(Accession: D3X7Z4), ermA (23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(B))(Accession: Q7DIC2), ant(9) )(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), NCTC11940_00344 (Protein of uncharacterized function (DUF3644))(Accession: A0A2X3P763), CSC83_13340 (Capsular biosynthesis protein)(Accession: A0A2840_00987) (Accession: A0A2S) Accession: A0A167QYC2), hmp (Flavohemoprotein) (Accession: A0A1D4MV18), SAP027A_009 (Antiseptic resistance protein) (Accession: D2J776), BN1321_390017 (Uncharacterized protein) (Accession: A0A0U1MRY5), , mecR1 (Methicillin resistance regulatory sensor-transducer MecR1) (Accession: A0A2Y2RBM2), cydA (Cytochrome d ubiquinol oxidase subunit I) (Accession: A0A167UFF7), lytS (Autolysis histidine kinase LytS) (01017PHage9), related protein) (Accession: A0A0U1MY60), SAMEA170867 4_03678 (Cell division inhibitor)(Accession: A0A2W2UEW9), capC (Tyrosine-protein phosphatase)(Accession: A0A0U1MX93), BTN44_14990 (Lipoprotein)(Accession: A0A0D3QB23), C0J46_00K500 (Ribok: rbs0A)(Accession: Abs0A)(Accession: Abs0A) (Accession: A0A2Y2LM71), ERS140147_00739 (Membrane protein) (Accession: A0A077VLJ7), bglA (6-phospho-beta-glucosidase) (Accession: A0A0D6GG52), pyrR (Bifunctional protein PyrR) (Accession: W8U6N8), vraR (DNA-binding), vraR (DNA-binding) response regulator)(Accession: D9IFM7), yxeP_2 (Carboxypeptidase, M20(D) family)(Accession: A0A0U1MEH9), orfX (Ribosomal RNA large subunit methyltransferase H)(Accession: A0A068W8S1), aroA (3-phosphoshikimate)(1-carboxyvinyltransferase)( Accession: A0A133PU31), queF (NADPH-dependent 7-cyano-7-deazaguanine reductase) (Accession: A0A077V3Q2), ydfH (Gluconate operon transcriptional repressor) (Accession: A0A1D4Q3C3), C7Q89_10110 (Abortive phage resistance protein) (Abortive phage resistance protein) , rpsC (30S ribosomal protein S3) (Accession: A0A1Q8DBU3), traP (Signal transduction protein TRAP)(Accession: A0A0U1N0V2), BSZ10_08125 (Alkaline-shock protein)(Accession: A0A1Q8DDB2), SAP059A_003 (Antiadhesin Pls, binding to squamous nasal epithelial cells)(Accession: D2) 2-oxobutanoate hydroxymethyltransferase) (Accession: A0A0Z1DQN8), rpsO (30S ribosomal protein S15) (Accession: A0A077VJU6), SAMEA1708674_01891 (Enterotoxin) (Accession: A0A1D4MQV2), ndk (Nucleoside) chain alcohol dehydrogenase)(Accession: A0A1D4IQF8), sbi (Immunoglobulin-binding protein sbi)(Accession: A0A0U1MSJ9), parB_2 (Chromosome partitioning protein ParB)(Accession: A0A0D3Q4U5), CSC83_07060 (Uncharacterized protein)_2 (Uncharacterized protein)_2 ( Ribosomal-protein-L7p-serine acetyltransferase)(Accession: A0A1D4MIM0), BTN44_15570 (Putative transposase)(Accession: B6V383), BN1321_380004 (DUF961 domain-containing protein)(Accession: A0A0U1MRA0), C7Q89_00130 (Protein-disulfide isomerase) (Protein-disulfide isomerase) ccession: A0A2W2RV36), truB_2 (Fibronectin-binding protein A)(Accession: A0A0U1MF71), ecsA_2 (Lantibiotic transport ATP-binding protein)(Accession: A0A0U1MEQ0), SAMEA1708674_03752 (Cell division protein (Putative))(Accession: A0A0U1MEQ0)(Cell division protein (Putative)) (Chaperone protein ClpB)(Accession: A0A2W3CJ21), argG (Argininosuccinate synthase)(Accession: A0A0D6WAS8), saeR (Response regulator SaeR)(Accession: A0A0U1MFP7), EP54_11640 (DegV domain-containing protein)6HNG6HNG0A0(Accession: A0A0D6WAS8) DNA-binding response regulator)(Accession: A0A2Y9TRS2), HMPREF3211_00240 (GntR family transcriptional regulator)(Accession: A0A133QB66), BSZ10_11980 (Transposon DNA-invertase)(Accession: A0A1Q8DBUN86), OX=1280 (Uncharacterized protein)(Accession: A0A2S1) , mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) and mnhE (Na (+) H (+) antiporter subunit E) (Accession: A0A077VW30) any one or more genes selected from the group consisting of or a protein encoded by the gene It is possible to provide a biomarker composition for determining the methicillin resistance or sensitive Staphylococcus aureus clone type.

The clone types are ST5 methicillin-resistant Staphylococcus aureus (ST5 MRSA), ST72 methicillin-resistant Staphylococcus aureus (ST72 MRSA), ST5 methicillin-sensitive Staphylococcus aureus (ST5 MSSA), and ST72 methicillin-sensitive Staphylococcus aureus (ST72 MSSA). ) may be selected from the group consisting of.

The present invention provides aacA (Aminoglycoside phosphotransferase APH(3'))(Accession: D3X7Z4), ermA (23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(B))(Accession: Q7DIC2), ant(9) )(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), NCTC11940_00344 (Protein of uncharacterized function (DUF3644))(Accession: A0A2X3P763), CSC83_13340 (Capsular biosynthesis protein)(Accession: A0A2840_00987) (Accession: A0A2S) Accession: A0A167QYC2), hmp (Flavohemoprotein) (Accession: A0A1D4MV18), SAP027A_009 (Antiseptic resistance protein) (Accession: D2J776), BN1321_390017 (Uncharacterized protein) (Accession: A0A0U1MRY5), , mecR1 (Methicillin resistance regulatory sensor-transducer MecR1) (Accession: A0A2Y2RBM2), cydA (Cytochrome d ubiquinol oxidase subunit I) (Accession: A0A167UFF7), lytS (Autolysis histidine kinase LytS) (01017PHage9), related protein) (Accession: A0A0U1MY60), SAMEA170867 4_03678 (Cell division inhibitor)(Accession: A0A2W2UEW9), capC (Tyrosine-protein phosphatase)(Accession: A0A0U1MX93), BTN44_14990 (Lipoprotein)(Accession: A0A0D3QB23), C0J46_00K500 (Ribok: rbs0A)(Accession: Abs0A)(Accession: Abs0A) (Accession: A0A2Y2LM71), ERS140147_00739 (Membrane protein) (Accession: A0A077VLJ7), bglA (6-phospho-beta-glucosidase) (Accession: A0A0D6GG52), pyrR (Bifunctional protein PyrR) (Accession: W8U6N8), vraR (DNA-binding), vraR (DNA-binding) response regulator)(Accession: D9IFM7), yxeP_2 (Carboxypeptidase, M20(D) family)(Accession: A0A0U1MEH9), orfX (Ribosomal RNA large subunit methyltransferase H)(Accession: A0A068W8S1), aroA (3-phosphoshikimate)(1-carboxyvinyltransferase)( Accession: A0A133PU31), queF (NADPH-dependent 7-cyano-7-deazaguanine reductase) (Accession: A0A077V3Q2), ydfH (Gluconate operon transcriptional repressor) (Accession: A0A1D4Q3C3), C7Q89_10110 (Abortive phage resistance protein) (Abortive phage resistance protein) , rpsC (30S ribosomal protein S3) (Accession: A0A1Q8DBU3), traP (Signal transduction protein TRAP)(Accession: A0A0U1N0V2), BSZ10_08125 (Alkaline-shock protein)(Accession: A0A1Q8DDB2), SAP059A_003 (Antiadhesin Pls, binding to squamous nasal epithelial cells)(Accession: D2) 2-oxobutanoate hydroxymethyltransferase) (Accession: A0A0Z1DQN8), rpsO (30S ribosomal protein S15) (Accession: A0A077VJU6), SAMEA1708674_01891 (Enterotoxin) (Accession: A0A1D4MQV2), ndk (Nucleoside) chain alcohol dehydrogenase)(Accession: A0A1D4IQF8), sbi (Immunoglobulin-binding protein sbi)(Accession: A0A0U1MSJ9), parB_2 (Chromosome partitioning protein ParB)(Accession: A0A0D3Q4U5), CSC83_07060 (Uncharacterized protein)_2 (Uncharacterized protein)_2 ( Ribosomal-protein-L7p-serine acetyltransferase)(Accession: A0A1D4MIM0), BTN44_15570 (Putative transposase)(Accession: B6V383), BN1321_380004 (DUF961 domain-containing protein)(Accession: A0A0U1MRA0), C7Q89_00130 (Protein-disulfide isomerase) (Protein-disulfide isomerase) ccession: A0A2W2RV36), truB_2 (Fibronectin-binding protein A)(Accession: A0A0U1MF71), ecsA_2 (Lantibiotic transport ATP-binding protein)(Accession: A0A0U1MEQ0), SAMEA1708674_03752 (Cell division protein (Putative))(Accession: A0A0U1MEQ0)(Cell division protein (Putative)) (Chaperone protein ClpB)(Accession: A0A2W3CJ21), argG (Argininosuccinate synthase)(Accession: A0A0D6WAS8), saeR (Response regulator SaeR)(Accession: A0A0U1MFP7), EP54_11640 (DegV domain-containing protein)6HNG6HNG0A0(Accession: A0A0D6WAS8) DNA-binding response regulator)(Accession: A0A2Y9TRS2), HMPREF3211_00240 (GntR family transcriptional regulator)(Accession: A0A133QB66), BSZ10_11980 (Transposon DNA-invertase)(Accession: A0A1Q8DBUN86), OX=1280 (Uncharacterized protein)(Accession: A0A2S1) , mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) and mnhE (Na(+) H(+) antiporter subunit E) (Accession: A0A077VW30) A probe that specifically binds to any one or more genes consisting of, amplifying the gene a primer, an antibody that specifically binds to the protein encoded by the gene, or a peptide having a binding domain specific to the protein It is possible to provide a kit for determining the methicillin-resistant or sensitive Staphylococcus aureus clone type comprising the.

The kit may be an RT-PCR kit, a DNA chip kit, or a protein chip kit, but is not limited thereto.

The present invention provides aacA (Aminoglycoside phosphotransferase APH(3'))(Accession: D3X7Z4), ermA (23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(B))(Accession : Q7DIC2), ant(9)(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), NCTC11940_00344 (Protein of uncharacterized function (DUF3644))(Accession: A0A2X3P763), CSC83_13340 (Capsular biosynthesis protein) (Capsular biosynthesis protein) (Capsular biosynthesis protein) ERS072840_00987 (Phage repressor protein)(Accession: A0A167QYC2), hmp (Flavohemoprotein)(Accession: A0A1D4MV18), SAP027A_009 (Antiseptic resistance protein)(Accession: D2J776), BN1321_390017 (Uncharacterized protein) (Uncharacterized protein) (Accession1) protein) (Accession: A0A2S6DEN9), mecR1 (Methicillin resistance regulatory sensor-transducer MecR1) (Accession: A0A2Y2RBM2), cydA (Cytochrome d ubiquinol oxidase subunit I) (Accession: A0A167UFF7), lytidine kinase (Accession: A0A167UFF7), lytidine LytS (AutolysisS kinase) ), SAMEA2445518_01017 (Hypothetical phage-related protein) (Accession: A0A 0U1MY60), SAMEA1708674_03678 (Cell division inhibitor)(Accession: A0A2W2UEW9), capC (Tyrosine-protein phosphatase)(Accession: A0A0U1MX93), BTN44_14990 (Lipoprotein)(Accession: A0A0D3QB23:bs), C0J r (Ribokinase) (Accession: A0A2Y2LM71), ERS140147_00739 (Membrane protein) (Accession: A0A077VLJ7), bglA (6-phospho-beta-glucosidase) (Accession: A0A0D6GG52), pyrR (Bifunctional protein PyrR) (Accession: W8U) DNA-binding response regulator) (Accession: D9IFM7), yxeP_2 (Carboxypeptidase, M20(D) family) (Accession: A0A0U1MEH9), orfX (Ribosomal RNA large subunit methyltransferase H) (Accession: A0A068W8S1), aroshikimate 1-phosphoshikimate carboxyvinyltransferase) (Accession: A0A133PU31), queF (NADPH-dependent 7-cyano-7-deazaguanine reductase) (Accession: A0A077V3Q2), ydfH (Gluconate operon transcriptional repressor) (Accession: A0A1DQ3C3), C7Q89 (AbortiveAccess4Q3C3), C7Q89 : B2Y834), rpsC (30S ribosomal protein S3) (Acc ession: A0A1Q8DBU3), traP (Signal transduction protein TRAP)(Accession: A0A0U1N0V2), BSZ10_08125 (Alkaline-shock protein)(Accession: A0A1Q8DDB2), SAP059A_003 (AntiadhesinJ Accessin2), nasal panthelial cells)(AntiadhesinJ Accession Pls, nasal panthelial cells) (3-methyl-2-oxobutanoate hydroxymethyltransferase)(Accession: A0A0Z1DQN8), rpsO (30S ribosomal protein S15)(Accession: A0A077VJU6), SAMEA1708674_01891 (Enterotoxin)(Accession: A0A1D4MQV2), ndk (Accession: A0A1D4MQV2), ndk (Accession: A0phosphate0 ndk) , SAMEA1708674_00628 (Short-chain alcohol dehydrogenase)(Accession: A0A1D4IQF8), sbi (Immunoglobulin-binding protein sbi)(Accession: A0A0U1MSJ9), parB_2 (Chromosome partitioning protein ParB)(Accession: A0A0D3Q4) (Uncharacterized protein) (UA0D3Q) A0A2L2EVY2), ydaF_2 (Ribosomal-protein-L7p-serine acetyltransferase)(Accession: A0A1D4MIM0), BTN44_15570 (Putative transposase)(Accession: B6V383), BN1321_380004 (DUF961d30), C7Q89U (Accession: A0A1D4MIM0) (Accession: A0A1D4MIM0) isulfide isomerase)(Accession: A0A2W2RV36), truB_2 (Fibronectin-binding protein A)(Accession: A0A0U1MF71), ecsA_2 (Lantibiotic transport ATP-binding protein)(Accession: A0A0U1MEQ0), SAMEA1708674_03752 (Cell division protein (Putative)) A0A1D3YL80), clpB (Chaperone protein ClpB)(Accession: A0A2W3CJ21), argG (Argininosuccinate synthase)(Accession: A0A0D6WAS8), saeR (Response regulator SaeR)(Accession: A0A0U1MFP7) (Accession: A0A0U1MFP7) NGV containing protein) ), RK64_10785 (DNA-binding response regulator)(Accession: A0A2Y9TRS2), HMPREF3211_00240 (GntR family transcriptional regulator)(Accession: A0A133QB66), BSZ10_11980 (Transposon DNA-invertase)(Accession: A0A1Q8DB86), OX=1280 Accession: A0A2S1FUN9), mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) and mnhE (Na(+) H(+) antiporter subunit E) (Accession: A0A077VW30) mRNA expression level of any one or more genes selected from the group consisting of checking the expression level of the protein encoded by the gene; and

It is possible to provide a method for determining the methicillin-resistant or sensitive Staphylococcus aureus clone type comprising the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control.

In more detail, the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control is aacA (Aminoglycoside phosphotransferase APH(3')) (Accession: D3X7Z4), ermA (23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(B))(Accession: Q7DIC2), ant(9)(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), NCTC11940_00344 (Protein of uncharacterized function (DUF3644))(Accession: A0A2X3P763), CSC83_13340 (Capsular biosynthesis protein)(Accession: A0A2S6DTY7), ERS072840_00987 (Phage repressor protein)(Accession: A0A167QYC2), hmp (Flavohemoprotein) protein) (Accession: D2J776), BN1321_390017 (Uncharacterized protein) (Accession: A0A0U1MRY5), CSC83_06955 (Phage major capsid protein) (Accession: A0A2S6DEN9), mecR1 (Methicillin resistance regulatory sensor-transducer MecR1) (Accession: Methicillin resistance regulatory sensor-transducer MecA0A2) Cytochrome d ubiquinol oxidase subunit I) (Accession: A0A167UFF7), lytS (Autolysis histidine kinase LytS) (Accession: A0A131JSM9), SAMEA244 5518_01017 (Hypothetical phage-related protein)(Accession: A0A0U1MY60), SAMEA1708674_03678 (Cell division inhibitor)(Accession: A0A2W2UEW9), capC (Tyrosine-protein phosphatase)(Accession: A0A0U1MX93), BTN346_1499000 (Accession: A0A0U1MX93), BTN044_1499000 (Transcriptional regulator) (Accession: A0A2W3FHQ4), rbsK (Ribokinase) (Accession: A0A2Y2LM71), and ERS140147_00739 (Membrane protein) (Accession: A0A077VLJ7) the mRNA expression level of any one or more genes selected from the group consisting of or encoded by the gene If the expression level of the protein is increased than that of the control, it may be determined as ST5 type methicillin-resistant Staphylococcus aureus (ST5 MRSA).

In addition, the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control is bglA (6-phospho-beta-glucosidase) (Accession: A0A0D6GG52), pyrR ( Bifunctional protein PyrR)(Accession: W8U6N8), vraR (DNA-binding response regulator)(Accession: D9IFM7), yxeP_2 (Carboxypeptidase, M20(D) family)(Accession: A0A0U1MEH9), orfX (Ribosomal RNA large subunit methyltransferase) ( Accession: A0A068W8S1), aroA (3-phosphoshikimate 1-carboxyvinyltransferase) (Accession: A0A133PU31), queF (NADPH-dependent 7-cyano-7-deazaguanine reductase) (Accession: A0A077V3Q2), ydfH (Gluconate operon transcriptional repressor) (Accession: A0A1D4Q3C3), C7Q89_10110 (Abortive phage resistance protein) (Accession: B2Y834) and rpsC (30S ribosomal protein S3) (Accession: A0A1Q8DBU3) mRNA expression level of any one or more genes selected from the group consisting of or If the expression level is increased than the control, it may be determined as ST72-type methicillin-resistant Staphylococcus aureus (ST5 MRSA).

In addition, the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control is traP (Signal transduction protein TRAP) (Accession: A0A0U1N0V2), BSZ10_08125 (Alkaline-shock) of a biological sample isolated from the sample. protein)(Accession: A0A1Q8DDB2), SAP059A_003 (Antiadhesin Pls, binding to squamous nasal epithelial cells)(Accession: D2JDZ0), panB (3-methyl-2-oxobutanoate hydroxymethyltransferase)(Accession: A0A0Z1DQN8), rpsom (Accession: A0A077VJU6), SAMEA1708674_01891 (Enterotoxin)(Accession: A0A1D4MQV2), ndk (Nucleoside diphosphate kinase)(Accession: A0A0U1MN06), SAMEA1708674_Q628 (Short-chain alcohol dehydrogenase) (Accession4) (Short-chain alcohol dehydrogenase) (Accession: A0A0U1MSJ9), parB_2 (Chromosome partitioning protein ParB)(Accession: A0A0D3Q4U5), CSC83_07060 (Uncharacterized protein)(Accession: A0A2L2EVY2), ydaF_2 (Ribosomal-protein-acetyltransfer4)_15 transposase) (Accession: B6V383), BN1321_380004 (DUF961 domain-con taining protein)(Accession: A0A0U1MRA0), C7Q89_00130 (Protein-disulfide isomerase)(Accession: A0A2W2RV36), truB_2 (Fibronectin-binding protein A)(Accession: A0A0U1MF71), ecsA0U1 (MEQ0) ATP-binding protein)(Accession: A0A0U1) (Lantibiotic transport) and SAMEA1708674_03752 (Cell division protein (Putative)) (Accession: A0A1D3YL80) When the mRNA expression level of any one or more genes selected from the group consisting of or the expression level of the protein encoding the gene is increased than the control group, ST5 type methicillin sensitive yellow Staphylococcus aureus (ST5 MSSA) may be determined.

In addition, the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control is clpB (Chaperone protein ClpB) (Accession: A0A2W3CJ21), argG (Argininosuccinate synthase) ( Accession: A0A0D6WAS8), saeR (Response regulator SaeR)(Accession: A0A0U1MFP7), EP54_11640 (DegV domain-containing protein)(Accession: A0A0D6HNG8), RK64_10785 (DNA-binding response regulator)(Accession: A0A2YMPREF3211) regulator) (Accession: A0A133QB66), BSZ10_11980 (Transposon DNA-invertase) (Accession: A0A1Q8DB86), OX=1280 (Uncharacterized protein) (Accession: A0A2S1FUN9), mprF (Phosphatidylglycerol lysyltransferase) (Accession: A077UML1) ) When the mRNA expression level of any one or more genes selected from the group consisting of H(+) antiporter subunit E) (Accession: A0A077VW30) or the expression level of the protein encoded by the gene is higher than that of the control group, ST72 type methicillin-sensitive staphylococcus aureus It may be determined as cocci (ST72 MSSA).

Methods for determining the mRNA expression level of the gene include RT-PCR, competitive RT-PCR (Competitive RT-PCR), real-time RT-PCR (Real-time RT-PCR), RNase protection assay (RPA; RNase protection assay), It may be to use any one selected from the group consisting of Northern blotting and a DNA chip, but is not limited thereto.

A method of confirming the expression level of the protein is Western blot, ELISA (enzyme linked immunosorbent asay), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immune diffusion method, locate ( rocket) immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, Complement Fixation Assay, FACS, mass spectrometry, and any one selected from the group consisting of a protein chip may be used. , but not limited thereto.

The biological sample of the present invention may be selected from the group consisting of blood, peripheral blood mononuclear cells (PBMC), sputum and urine, but is not limited thereto.

A "primer" of the present invention is a nucleic acid sequence having a short free 3' hydroxyl group, a short nucleic acid sequence capable of base pairing with a complementary template and serving as a starting point for template strand copying. say The primer is capable of initiating DNA synthesis in the presence of a reagent for polymerization (ie, DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. PCR conditions and lengths of sense and antisense primers may be appropriately selected according to techniques known in the art.

"Probe" of the present invention refers to a nucleic acid fragment such as RNA or DNA corresponding to several bases to several hundred bases in length that can specifically bind to other than mRNA, and is labeled to determine the presence or absence of a specific mRNA and the amount of expression. can be checked The probe may be manufactured in the form of an oligonucleotide probe, a single-stranded DNA probe, a double-stranded DNA probe, an RNA probe, or the like. Suitable probe selection and hybridization conditions can be appropriately selected according to techniques known in the art.

"Antibody" of the present invention is a term known in the art and refers to a specific immunoglobulin directed against an antigenic site. Those prepared through injection of one or more proteins mentioned above or those purchased commercially may be used. In addition, the antibody includes polyclonal antibodies, monoclonal antibodies, and fragments capable of binding to an epitope.

"Peptide" of the present invention refers to a polypeptide that does not have the structure of an intact antibody, but has a specific antigen binding site (binding domain) directed to the antigenic site. Such peptides include functional fragments of antibody molecules that are not intact antibodies having two light chains and two heavy chains. A functional fragment of an antibody molecule means a fragment having at least an antigen-binding function. The peptide comprises at least 7 amino acids, preferably at least 9 amino acids, more preferably at least 12 amino acids.

Hereinafter, to help the understanding of the present invention, examples will be described in detail. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Example 1> Protein discovery

Blood was collected from a total of 40 patients infected with MSSA and MRSA with ST5 genotype, MSSA and MRSA with ST72 genotype, and Staphylococcus aureus was isolated and cultured, and after dissolving the cells, BCA array was performed. Thus, 100 μg of each pooled sample set was prepared. Details of each sample group are as follows.

1) ST5 MSSA: 10 types in total (8 arg fx + 2 arg dfx)

2) ST5 MRSA: 10 types in total (1 arg fx + 4 arg dfx + 5 arg dfx/hVISA moderators)

3) ST72 MSSA: 10 types in total (arg fx 10 persons)

4) ST72 MRSA: Total 10 types (arg fx/hVISA 1 person + arg fx 7 person + arg dfx/hVISA 1 person + arg dfx 1 person)

Proteins extracted from each sample, a 30 kDa cutoff-filter unit was used as a digestion unit FASP (filter-aided sample preparation) technique using a peptide was prepared.

Specifically, 100 μl of 8 M urea containing a protease / phosphatase inhibitor cocktail was mixed with a concentrated small protein fraction, and denatured by centrifugation at 13,000 rpm for 10 minutes ( denaturation). Thereafter, disulfide bonds were reduced at 37° C. using 50 mM dithiothreitol, and the reduction was performed by centrifugation at 13,000 rpm for 10 minutes. After performing an alkylation reaction under light blocking conditions using 55 mM iodoacetamide, centrifugation was performed at 13,000 rpm for 10 minutes. Then, 200 ml of 50 mM ammonium bicarbonate was added and centrifuged at 13,000 rpm for 10 minutes.

LysS/trypsin mixed protease 100 ng was treated in each sample, and digestion was performed overnight at 37 ℃. Thereafter, centrifugation was performed at 13,000 rpm for 10 minutes, and after stopping the reaction with 10% formic acid, desalting was performed using a C18 reverse phase desalting cartridge.

The desalted peptide was dried using SpeedVac and then dissolved in 0.1% formic acid to perform LC-MS analysis. Q-Exactive Plus BioPharm version (Thermo, USA) was used for MS equipment, and UltiMate™3000 RSLCnano System (Thermo, USA) was used for LC equipment. Analysis was performed with a coupled ion trap MS.

LC was separated for a total of 150 minutes, including a concentration gradient of 120 minutes between Solution A (5% acetonitrile, 0.1% formic acid) and solution B (95% acetonitrile, 0.1% formic acid), and the analytical column used was AQUA C18 ( The peptide mixture was separated using a fused silica capillary column with an inner diameter of 75 µm, an outer diameter of 360 µm, and a length of 50 cm filled with 5 µm, 125A). The separated peptide was ion-sprayed using an electrospray ionization (ESI) voltage of 2000 V, and the ion emitter was subjected to mass spectrometry through a tapered ion spray emitter with an inner diameter of 20 μm. Spectral data was obtained by injecting into

Orbitrap MS analysis was performed 8 times using HCD (higher energy collision dissociation, 25% energy level) method and Orbitrap MS after performing one survey scan (resolution 30,000) with ion trap MS in the range of 400 to 2000 m/z. MS/MS (resolution 60,000) analysis was performed. The detection of overlapping peptide ions was minimized through the dynamic exclusion option of 5 min. The auto gain control target setting of the ion trap was 3E04 for Full MS and 2E5 for FT MS/MS.

The acquired RAW data were subjected to qualitative analysis and label-free quantitative analysis using Proteome Discoverer (Thermo, USA, version 2.2), a database analysis software based on Sequest algorithm. Peptide modifications were performed with fixed modifications by cysteine carbamidomethylation, methionine oxidation, and deamidation. For trypsin uncleaved trypsin misscleavage), a value of “1” was used, the latest swissprot sequence DB was used for the sequence database, and quantitative analysis using peptide ion strength was performed for label-free quantitation.

As a result of duplication of qualitative and label-free quantitative analysis for the four strain groups by the above process, 1383 proteins were identified in the MSSA group and 1399 proteins in the ST5 MRSA group as shown in FIG. 2 . 1374 proteins were identified in ST72-type MSSA and 1368 proteins were identified in ST72 MRSA.

<Example 2> Protein screening

The accession code was unified with the gene names of 1,361 proteins commonly identified in ST5 MSSA and MRSA and 1,337 proteins commonly identified in ST72 MSSA and MRSA identified in the previous experiment. The abundance of each protein was extracted, and proteins in which abundance did not exist or were not identified in common were excluded.

ANOVA test was performed on 8 data sets, which are the duplication results for 4 groups, and hierarchical cluster analysis was performed by selecting 164 proteins showing a significance of 0.01 or less based on the Benjamini-Hochberg FDR.

As a result, as shown in FIG. 3, four groups of ST5 MRSA, ST5 MSSA, ST72 MRSA, and ST72 MSSA were classified into nine clusters of proteins showing the same quantitative expression pattern in eight data sets that were analyzed twice.

In addition, among the nine clusters, four clusters of FIGS. 4 to 7 in which the expression levels were consistently increased in the ST5 MRSA, ST5 MSSA, ST72 MRSA and ST72 MSSA groups were finally selected.

The four selected clusters not only serve the purpose of classifying MRSA and MSSA, but can also be utilized to confirm whether nosocomial infection (ST5) or local infection (ST72). In addition, it can be used for rapid diagnosis by detecting each group-specific protein present in a patient's appropriate sample (blood, PBMC, sputum or urine, etc.) have.

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (11)

delete delete ant(9)(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), CSC83_13340 (Capsular biosynthesis protein)(Accession: A0A2S6DTY7), SAP027A_009 (Antiseptic resistance protein)(Accession: D2J776), CSC83_06955 (Phage major capsid protein) Accession: A0A2S6DEN9), yxeP_2 (Carboxypeptidase, M20(D) family)(Accession: A0A0U1MEH9), aroA (3-phosphoshikimate 1-carboxyvinyltransferase)(Accession: A0A133PU31), SAP059A_003 (Antiadhesin nasal) (Accession epithelial cells, binding to squamous Pls, : D2JDZ0), ndk (Nucleoside diphosphate kinase)(Accession: A0A0U1MN06), CSC83_07060 (Uncharacterized protein)(Accession: A0A2L2EVY2), HMPREF3211_00240 (GntRBS family transcriptional regulator)(AccessTransposion: A0A133QB66) (Accession: A0A133QB66) (Accession: A0A133QB66) A0A1Q8DB86), and a probe that specifically binds to mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) gene, a primer for amplifying the gene, an antibody that specifically binds to a protein encoded by the gene, or an antibody specific for the protein ST5 methicillin-resistant Staphylococcus aureus (ST5 MRSA), ST72 methicillin-resistant Staphylococcus aureus comprising a peptide having a binding domain Bacillus (ST72 MRSA), ST5 methicillin-sensitive Staphylococcus aureus (ST5 MSSA), and ST72 methicillin-sensitive Staphylococcus aureus (ST72 MSSA) methicillin-resistant or sensitive Staphylococcus aureus clonetype identification kit. 4. The kit according to claim 3, wherein the kit is an RT-PCR kit, a DNA chip kit, or a protein chip kit. ant(9)(Streptomycin 3''-adenylyltransferase)(Accession: Q7DIC3), CSC83_13340 (Capsular biosynthesis protein)(Accession: A0A2S6DTY7), SAP027A_009 (Antiseptic resistance protein) (Accession: D2J776), CSC83_06955 (Phage major capsid protein)(Accession: A0A2S6DEN9), yxeP_2 (Carboxypeptidase, M20(D) family)(Accession: A0A0U1MEH9), aroA (3-phosphoshikimate 1-carboxyvinyltransferase)(Accession: A0A133PU31), SAP059A_003 (Antiadhesin Pls, binding to squamous nasal epithelial cells)(Accession: D2JDZ0), ndk (Nucleoside diphosphate kinase)(Accession: A0A0U1MN06), CSC83_07060 (Uncharacterized protein)(Accession: A0A2L2EVY2), HMPREF3211_00240_11) (GntR family transcriptionon: A0A2EVY2), (GntR family transcriptionon regulator) DNA-invertase) (Accession: A0A1Q8DB86), and mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) determining the mRNA expression level of the gene or the expression level of the protein encoded by the gene; and
ST5 methicillin-resistant Staphylococcus aureus (ST5 MRSA), ST72 methicillin-resistant Staphylococcus aureus, comprising the step of determining whether the expression is increased by comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control. A method for determining a methicillin-resistant or sensitive Staphylococcus aureus clonetype selected from the group consisting of cocci (ST72 MRSA), ST5 methicillin-sensitive Staphylococcus aureus (ST5 MSSA), and ST72 methicillin-sensitive Staphylococcus aureus (ST72 MSSA). According to claim 5, wherein the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control is ant(9) (Streptomycin 3''-adenylyltransferase) (Accession) of a biological sample isolated from a specimen : Q7DIC3), CSC83_13340 (Capsular biosynthesis protein) (Accession: A0A2S6DTY7), SAP027A_009 (Antiseptic resistance protein) (Accession: D2J776), and CSC83_06955 (Phage major capsid protein) (Accession: any one selected from the group consisting of A0A2S6DEN9) or more When the mRNA expression level of the gene or the expression level of the protein encoded by the gene is increased than the control, methicillin-resistant or sensitive Staphylococcus aureus clone type discrimination, characterized in that it is determined as ST5 type methicillin-resistant Staphylococcus aureus (ST5 MRSA) Way. According to claim 5, wherein the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control is yxeP_2 (Carboxypeptidase, M20 (D) family) (Accession: A0A0U1MEH9 of a biological sample isolated from a specimen) ), and aroA (3-phosphoshikimate 1-carboxyvinyltransferase) (Accession: A0A133PU31) When the mRNA expression level of any one or more genes selected from the group or the expression level of the protein encoded by the gene is increased than the control group, ST72 type methicillin A method for determining a methicillin-resistant or sensitive Staphylococcus aureus clone type, characterized in that it is determined as resistant Staphylococcus aureus (ST5 MRSA). According to claim 5, wherein the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control is SAP059A_003 (Antiadhesin Pls, binding to squamous nasal epithelial cells) (Accession) of the biological sample isolated from the specimen. : D2JDZ0), ndk (Nucleoside diphosphate kinase) (Accession: A0A0U1MN06), and CSC83_07060 (Uncharacterized protein) (Accession: A0A2L2EVY2) mRNA expression level of any one or more genes selected from the group consisting of or expression of a protein encoded by the gene When the level is increased than the control, methicillin-resistant or sensitive Staphylococcus aureus clone type discrimination method, characterized in that it is determined as ST5 type methicillin-sensitive Staphylococcus aureus (ST5 MSSA). The method according to claim 5, wherein the step of comparing the mRNA expression level of the gene or the expression level of the protein encoded by the gene with a control comprises HMPREF3211_00240 (GntR family transcriptional regulator) (Accession: A0A133QB66), BSZ10_11980 of a biological sample isolated from a specimen. (Transposon DNA-invertase) (Accession: A0A1Q8DB86), and mprF (Phosphatidylglycerol lysyltransferase) (Accession: A0A077UML1) The mRNA expression level of any one or more genes selected from the group or the expression level of the protein encoded by the gene is higher than that of the control group. Methicillin-resistant or sensitive Staphylococcus aureus clone type discrimination method, characterized in that when it is increased, it is determined as ST72-type methicillin-sensitive Staphylococcus aureus (ST72 MSSA). According to claim 5, wherein the method for determining the mRNA expression level of the gene is RT-PCR, competitive RT-PCR (Competitive RT-PCR), real-time RT-PCR (Real-time RT-PCR), RNase protection assay (RPA) ; RNase protection assay), Northern blotting and methicillin-resistant or sensitive Staphylococcus aureus clone type identification method, characterized in that using any one selected from the group consisting of a DNA chip. According to claim 5, wherein the method for determining the expression level of the protein is Western blot, ELISA (enzyme linked immunosorbent asay), radioimmunoassay (RIA), radioimmunodiffusion method (radioimmunodiffusion), Ouchterlony (Ouchterlony) Any one selected from the group consisting of immune diffusion method, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, Complement Fixation Assay, FACS, mass spectrometry and protein chip Method for determining the methicillin-resistant or sensitive Staphylococcus aureus clone type, characterized in that using one.

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