PCR primer set for detection of carbapenemase-producing enterobacterales and uses thereof

Abstract

The present invention provides a novel means for detecting various variants of carbapenemase-producing Enterobacterales (CPE) in which carbapenemase is an enzyme breaking down carbapenem antibiotics. The primer sets of the present invention are capable of detecting, with high accuracy in silico, CPE-associated genes such as KPC, NDM, OXA-48-family, GES, VIM-2-like, IMP G1, and variants thereof. Even when the primer sets are combined for multiplex polymerase chain reaction (PCR), nonspecific bands do not appear at the same annealing temperature, with the generation of PCR amplification products of different sizes, allowing detection of various CPE-related genes and variants thereof in a single PCR reaction. Furthermore, the use of probes that specifically bind to each PCR amplification product enables application in real-time PCR, thus providing a highly accurate and specific means for detecting carbapenemase producing Enterobacterales (CPE) strains.

Description

PCR primer set for detection of carbapenemase-producing Enterobacteriaceae and use thereof

[0001] The present invention relates to a PCR primer set for detecting carbapenemase producing Enterobacterales (CPE) and its use.

[0002] Enterobacterales is an order of bacteria belonging to the class Gammaproteobacteria. Bacteria belonging to this genus are characterized by being gram-negative, non-spore-forming, facultatively anaerobic, and rod-shaped. Bacteria belonging to Enterobacterales are recognized as a major cause of community-acquired and healthcare-associated infections, and many strains of Enterobacterales are found to be ESBL (extended spectrum β-lactamase)-positive multidrug-resistant Enterobacteria, drawing attention as a serious healthcare problem. Carbapenems are currently used as antibiotics to treat ESBL-producing Enterobacteriaceae infections. However, with the increased use of carbapenems, the number of carbapenem-resistant Enterobacterales (CRE) or carbapenemase-producing Enterobacterales (CPE) strains has been reported to increase. The rapid increase in CPE strains has emerged as a serious global public health problem, and the detection of CPE strains and examination of related resistance genes for the purpose of infection epidemiology, appropriate diagnosis, and treatment are important tasks for solving infectious diseases and public health problems caused by CRE or CPE strains.

[0003] Carbapenemase, or carbapenemase, produced by the CPE strain is an enzyme that degrades carbapenem antibiotics. Carbapenemases can recognize and degrade beta-lactam antibiotics, and exhibit resistance to many types of beta-lactamase inhibitors. Carbapenemase enzymes are classified into class A, class B, class C, and class D based on the similarity of their amino acid sequences. Class A includes the KPC and GES genes, class B includes the VIM, IMP, and NDM genes, and class D includes the OXA gene. Each gene has a wide variety of genotypes through subtle mutations. As of March 2024, 194 variants have been reported for KPC, 60 for NDM, 1,182 for OXA, 59 for GES, 84 for VIM, and 98 for IMP. Due to these genetic characteristics, existing molecular genetic methods have limitations in identifying newly reported genotypes.

[0004] From June 2017 to December 2020, the number of domestic human-derived CPE isolated through the National Institute of Environmental Research was 24,188. The isolation frequency by genotype was the highest for KPC-2 at 73.82%, followed by NDM-1 (12.93%), NDM-5 (4.57%), OXA-181 (1.81%), and KPC-4 (0.68%). In the genotype distribution of 18,295 human-derived CPE cases investigated in 10 university hospitals in Korea in 2021, KPC was the most common at 80.89%, followed by NDM (14.51%) and OXA (1.92%). In addition to KPC, NDM, and OXA, which occur frequently in Korea, GES, VIM, and IMP also occur sporadically, although less frequently. Therefore, a broad detection method for the six genotypes is required.

[0005] [Prior Art Literature]

[0006] [Patent Document]

[0007] Korean Patent Publication No. 10-2012-0039842 (Published on April 26, 2012)

[0008] The purpose of the present invention is to provide a new means for detecting various mutations of carbapenemase-producing Enterobacterales (CPE) strains, which are enzymes that degrade carbapenems, which are antibiotics.

[0009] The present invention relates to a first primer set comprising a forward primer represented by SEQ ID NO: 1 or SEQ ID NO: 3 and a reverse primer represented by SEQ ID NO: 2 or SEQ ID NO: 4; a second primer set comprising a forward primer represented by SEQ ID NO: 5 or SEQ ID NO: 7 and a reverse primer represented by SEQ ID NO: 6 or SEQ ID NO: 8; a third primer set comprising a forward primer represented by SEQ ID NO: 9 or SEQ ID NO: 11 and a reverse primer represented by SEQ ID NO: 10 or SEQ ID NO: 12; a fourth primer set comprising a forward primer represented by SEQ ID NO: 13 or SEQ ID NO: 15 and a reverse primer represented by SEQ ID NO: 14 or SEQ ID NO: 16; a fifth primer set comprising a forward primer represented by SEQ ID NO: 17 or SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 18 or SEQ ID NO: 20; The present invention provides a composition for detecting carbapenemase producing Enterobacterales (CPE), comprising at least one primer set selected from the group consisting of a forward primer represented by SEQ ID NO: 21 or SEQ ID NO: 23 and a sixth primer set consisting of a reverse primer represented by SEQ ID NO: 22 or SEQ ID NO: 24.

[0010] In addition, the present invention comprises a step of isolating DNA from a sample (step 1); a step of performing a polymerase chain reaction (PCR) using at least one primer set selected from the group consisting of a first primer set comprising a forward primer represented by SEQ ID NO: 1 and a reverse primer represented by SEQ ID NO: 2, using the DNA separated in the first step as a template, a second primer set comprising a forward primer represented by SEQ ID NO: 5 and a reverse primer represented by SEQ ID NO: 6, a third primer set comprising a forward primer represented by SEQ ID NO: 9 and a reverse primer represented by SEQ ID NO: 10, a fourth primer set comprising a forward primer represented by SEQ ID NO: 13 and a reverse primer represented by SEQ ID NO: 16, a fifth primer set comprising a forward primer represented by SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 20, and a sixth primer set comprising a forward primer represented by SEQ ID NO: 21 and a reverse primer represented by SEQ ID NO: 22; a step of performing electrophoresis on a PCR amplification product obtained in the second step (step 3); And a step (fourth step) of detecting a carbapenemase-producing Enterobacterales (CPE) strain according to the presence or absence of a PCR amplification product confirmed in the third step is provided. The present invention provides a method for detecting a carbapenemase-producing Enterobacterales (CPE) strain.

[0011] According to the present invention, the primer sets of the present invention detect CPE-related genes, KPC, NDM, OXA-48-family, GES, VIM-2-like, IMP G1 and their variants in silico with high accuracy, and when the primer sets are combined to perform multiplex PCR, no non-specific band appears even at the same annealing temperature, and since each PCR amplification product having a different size is formed, various CPE-related genes and their variants can be detected with a single PCR, and since it can be applied to real-time PCR using probes that specifically bind to each PCR amplification product, it can be provided as a means with high accuracy and specificity for detecting carbapenemase-producing Enterobacterales (CPE) strains.

[0012] Figure 1 shows the results of gradient PCR according to the composition of a primer set for detecting carbapenemase-producing Enterobacterales (CPE) according to the present invention.

[0013] Figure 2 shows the results of Triplex PCR for detecting carbapenemase-producing Enterobacterales (CPE) according to the present invention.

[0014] Figure 3 is an amplification plot showing real-time PCR results according to probe and template concentration for detection of carbapenemase-producing Enterobacterales (CPE) according to the present invention.

[0015] Figure 4 is a Triplex real-time PCR amplification plot according to the number of Step 1 cycles of the Triplex PCR set for detecting carbapenemase-producing Enterobacterales (CPE) according to the present invention.

[0016] FIG. 5 is a Triplex real-time PCR amplification plot showing the detection limit of a Triplex PCR set for detecting carbapenemase-producing Enterobacterales (CPE) according to the present invention.

[0017] Figure 6 shows the results of the IMP G1 primer set1 in silico test according to the phylogenetic cluster of the IMP gene. Red circles (undetectable), blue squares (detectable).

[0018] The terms used in this specification have been selected from widely used, current terms, taking into account the functions of the present invention. However, these terms may vary depending on the intentions of those skilled in the art, precedents, the emergence of new technologies, etc. Furthermore, in certain cases, terms may be arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in this invention should not be defined simply as names, but rather based on their inherent meanings and the overall content of the present invention.

[0019] Unless otherwise defined, 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 this invention pertains. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0020] Hereinafter, the present invention will be described in more detail.

[0021] The present invention provides a new means for detecting various mutations in carbapenemase-producing Enterobacterales (CPE) strains, which are enzymes that degrade carbapenems, which are antibiotics. In order to do this, a primer set capable of comprehensively detecting various genes related to CPE and their mutations was designed, and a primer set capable of detecting mutations in each gene was selected in silico. The identified primer sets detect CPE-related genes and their mutations with high accuracy, and since non-specific bands do not appear even at the same annealing temperature and individual PCR amplification products of different sizes are formed, various CPE-related genes and their mutations can be detected in a single PCR. Based on this, the present invention aims to provide a means for detecting various CPE-related genes and their mutations in a single PCR.

[0022] The present invention relates to a first primer set comprising a forward primer represented by SEQ ID NO: 1 or SEQ ID NO: 3 and a reverse primer represented by SEQ ID NO: 2 or SEQ ID NO: 4; a second primer set comprising a forward primer represented by SEQ ID NO: 5 or SEQ ID NO: 7 and a reverse primer represented by SEQ ID NO: 6 or SEQ ID NO: 8; a third primer set comprising a forward primer represented by SEQ ID NO: 9 or SEQ ID NO: 11 and a reverse primer represented by SEQ ID NO: 10 or SEQ ID NO: 12; a fourth primer set comprising a forward primer represented by SEQ ID NO: 13 or SEQ ID NO: 15 and a reverse primer represented by SEQ ID NO: 14 or SEQ ID NO: 16; a fifth primer set comprising a forward primer represented by SEQ ID NO: 17 or SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 18 or SEQ ID NO: 20; The present invention provides a composition for detecting carbapenemase producing Enterobacterales (CPE), comprising at least one primer set selected from the group consisting of a forward primer represented by SEQ ID NO: 21 or SEQ ID NO: 23 and a sixth primer set consisting of a reverse primer represented by SEQ ID NO: 22 or SEQ ID NO: 24.

[0023] The first primer set can specifically detect a KPC gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 46 to SEQ ID NO: 120. When the first primer set is a forward primer represented by SEQ ID NO: 1 and a reverse primer represented by SEQ ID NO: 2 (KPC primer set1), a KPC gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 46 to SEQ ID NO: 55 is specifically detected, and when the first primer set is a forward primer represented by SEQ ID NO: 3 and a reverse primer represented by SEQ ID NO: 4 (KPC primer set2), a KPC gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 56 to SEQ ID NO: 120 is specifically detected.

[0024] Specifically, the above KPC gene(NCBI GenBank No. NG_049253.1) or this mutation (variant) is specifically detected, and the above KPC gene mutation is NCBI GenBank No. NG_049243.1, NG_049244.1, NG_049245.1, NG_049246.1, NG_049247.1, NG_049248.1, NG_049249.1, NG_049250.1, NG_049251.1, NG_049252.1, NG_049253.1, NG_049254.1, NG_049255.1, NG_049256.1, NG_049257.1, NG_049258.1, NG_049259.1, NG_049260.1, NG_049261.1, NG_049262.1, NG_051167.1, NG_051469.1, NG_052581.1, NG_052862.1, NG_054685.1, NG_055494.1, NG_055495.1, NG_055580.1, NG_056170.1, NG_057447.1, NG_060524.1, NG_060569.1, NG_061389.1, NG_061612.1, NG_062357.1, NG_063841.1, NG_064726.1, NG_064727.1, NG_064728.1, NG_065427.1, NG_065876.1, NG_065877.1, NG_065878.1, NG_067224.1, NG_067225.1, NG_067226.1, NG_068016.1, NG_068176.1, NG_068177.1, NG_068507.1, NG_068508.1, NG_070177.1, NG_070178.1, NG_070179.1, NG_070180.1, NG_070739.1, NG_070740.1, NG_070741.1, NG_070742.1, NG_070743.1, NG_070895.1, NG_070896.1, NG_070897.1, NG_071203.1, NG_071204.1, NG_071205.1, NG_073465.1, NG_073466.1, NG_073467.1, NG_073468.1, NG_073469.1, NG_073470.1, NG_073471.1, NG_074714.1, NG_074715.1, NG_074716.1, NG_074717.1, NG_074718.1, NG_074719.1, NG_074720.1, NG_074721.1, NG_074722.1, NG_074723.1, NG_074724.1, NG_076666.1, NG_076667.1, NG_076680.1, NG_076681.1, NG_078032.1, NG_078037.1, NG_078038.1, NG_078051.1, NG_078052.1, NG_078053.1, NG_078054.1, NG_078055.1, NG_078056.1, NG_078063.1, NG_079230.1, NG_079231.1, NG_079232.1, NG_079233.1, NG_079888.1, NG_079889.1, NG_079890.1, NG_079891.1, NG_079892.1, NG_079893.1, NG_079894.1, NG_079895.1, NG_079896.1, NG_079897.1, NG_079898.1, NG_080778.1, NG_080779.1, NG_080780.1, NG_081070.1, NG_081071.1, NG_081072.1, NG_081699.1, NG_081700.1, NG_081783.1, NG_081784.1, NG_081791.1, NG_088394.1, NG_088395.1, NG_088396.1, NG_088397.1, NG_088398.1, NG_088399.1, NG_088400.1, NG_088401.1, NG_088402.1, NG_088403.1, NG_088404.1, NG_088405.1, NG_148622.1, NG_148623.1, NG_148624.1, NG_148625.1, NG_148626.1, NG_148627.1, NG_149659.1, NG_149660.1, NG_149661.1, NG_149662.1, NG_157007.1, NG_157008.1, NG_157009.1, NG_157010.1, NG_157011.1, NG_157012.1, NG_157013.1, NG_157014.1, NG_157015.1, NG_157016.1, NG_203393.1, NG_203394.1, NG_203395.1, NG_203396.1, NG_203397.1, NG_228670.1, NG_228671.1, NG_228672.1, NG_228673.1, NG_228674.1, NG_231545.1, NG_231546.1, NG_231547.1, NG_231548.1, NG_231549.1, NG_231550.1, NG_231551.1, NG_231552.1, NG_231553.1, NG_242182.1, NG_242183.1, NG_242184.1, NG_242185.1, NG_242186.1, NG_242187.1, NG_242188.1, NG_242189.1, NG_242190.1, NG_242293.1, NG_242294.1, NG_242295.1, NG_242296.1, NG_242297.1, NG_242298.1, NG_242299.1, NG_242300.1, NG_242301.1, and NG_242302.1 are the selected ones.

[0025] The second primer set can specifically detect an NDM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 121 to SEQ ID NO: 143. When the second primer set is a forward primer represented by SEQ ID NO: 5 and a reverse primer represented by SEQ ID NO: 6 (NDM primer set1), an NDM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 121 to SEQ ID NO: 139 is specifically detected, and when the second primer set is a forward primer represented by SEQ ID NO: 7 and a reverse primer represented by SEQ ID NO: 8 (NDM primer set2), an NDM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 140 to SEQ ID NO: 143 is specifically detected.

[0026] Specifically, the above 2nd primer set specifically detects the NDM gene (NCBI GenBank No. NG_049326.1) or this variant, and the above NDM gene variant is NCBI GenBank No. NG_049326.1, NG_049327.1, NG_049328.1, NG_049329.1, NG_049330.1, NG_049331.1, NG_049332.1, NG_049333.1, NG_049334.1, NG_049335.1, NG_049336.1, NG_049337.1, NG_049338.1, NG_049339.1, NG_049340.1, NG_049341.1, NG_052662.1, NG_052866.1, NG_055498.1, NG_055664.1, NG_057455.1, NG_057612.1, NG_060570.1, NG_060571.1, NG_062358.1, NG_064729.1, NG_066711.1, NG_067144.1, NG_067145.1, NG_071206.1, NG_071207.1, NG_074726.1, NG_076641.1, NG_076642.1, NG_076661.1, NG_076662.1, NG_076664.1, NG_076842.1, NG_076843.1, Any one selected from NG_078034.1, NG_080782.1, NG_080783.1, NG_081701.1, NG_088409.1, NG_148636.1, NG_148637.1, NG_148638.1, NG_148639.1, NG_149663.1, NG_149664.1, NG_157017.1, NG_157018.1, NG_157019.1, NG_157020.1, NG_157021.1, NG_203399.1, NG_203400.1, NG_203401.1, NG_203402.1, and NG_231554.1.

[0027] The third primer set can specifically detect an OXA-48 gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 144 to SEQ ID NO: 183. When the third primer set is a forward primer represented by SEQ ID NO: 9 and a reverse primer represented by SEQ ID NO: 10 (OXA-48 primer set1), an OXA-48 gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 144 to SEQ ID NO: 146 is specifically detected, and when the third primer set is a forward primer represented by SEQ ID NO: 11 and a reverse primer represented by SEQ ID NO: 12 (OXA-48 primer set2), an OXA-48 gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 147 to SEQ ID NO: 183 is specifically detected.

[0028] Specifically, the above-mentioned 제3 primer set will specifically detect the OXA-48-family gene (NCBI GenBank No. NG_049762.1) or its mutation(variant), and the above-mentioned OXA-48-family gene mutation is NCBI GenBank No. NG_049461.1, NG_049462.1, NG_049482.1, NG_049495.1, NG_049502.1, NG_049528.1, NG_049539.1, NG_049540.1, NG_049542.1, NG_049661.1, NG_049694.1, NG_049704.1, NG_049722.1, NG_049724.1, NG_049725.1, NG_049762.1, NG_049766.1, NG_049783.1, NG_049794.1, NG_052051.1, NG_054666.1, NG_054693.1, NG_054959.1, NG_055475.1, NG_055476.1, NG_055490.1, NG_055499.1, NG_055666.1, NG_056053.1, NG_061613.1, NG_064749.1, NG_064751.1, NG_065443.1, NG_066509.1, NG_066755.1, NG_070182.1, NG_070183.1, NG_070185.1, Any one selected from NG_070186.1, NG_070747.1, NG_070750.1, NG_077983.1, NG_077984.1, NG_078041.1, NG_078042.1, NG_079243.1, NG_081705.1, NG_088410.1, NG_149665.1, NG_203403.1, NG_203404.1, NG_203405.1, NG_203408.1, NG_203409.1, NG_228675.1, NG_228676.1, NG_228677.1, and NG_242303.1.

[0029] The above fourth primer set can specifically detect a GES gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 184 to SEQ ID NO: 217. When the fourth primer set is a forward primer represented by SEQ ID NO: 13 and a reverse primer represented by SEQ ID NO: 14 (GES primer set1) or a forward primer represented by SEQ ID NO: 13 and a reverse primer represented by SEQ ID NO: 16 (GES primer mix1), a GES gene or a variant thereof comprising any one of the base sequences of SEQ ID NOs: 184 to 208 is specifically detected, and when the fourth primer set is a forward primer represented by SEQ ID NO: 15 and a reverse primer represented by SEQ ID NO: 16 (GES primer set2) or a forward primer represented by SEQ ID NO: 15 and a reverse primer represented by SEQ ID NO: 14 (GES primer mix2), a GES gene or a variant thereof comprising any one of the base sequences of SEQ ID NOs: 209 to 217 is specifically detected.

[0030] Specifically, the above-mentioned GES gene (NCBI GenBank No. NG_049111.1) or this variant is specifically detected, and the above-mentioned GES gene is NCBI GenBank No. NG_049111.1, NG_049112.1, NG_049113.1, NG_049114.1, NG_049115.1, NG_049116.1, NG_049117.1, NG_049118.1, NG_049119.1, NG_049120.1, NG_049121.1, NG_049122.1, NG_049123.1, NG_049124.1, NG_049125.1, NG_049126.1, NG_049127.1, NG_049128.1, NG_049129.1, NG_049130.1, NG_049131.1, NG_049132.1, NG_049133.1, NG_049134.1, NG_049135.1, NG_049136.1, NG_049137.1, NG_049138.1, NG_049139.1, NG_049140.1, NG_049141.1, NG_051512.1, NG_054709.1, NG_056177.1, NG_060520.1, NG_060521.1, NG_060522.1, NG_062215.1, NG_062216.1, Any one selected from NG_062356.1, NG_065425.1, NG_065870.1, NG_065871.1, NG_070735.1, NG_070736.1, NG_071202.1, NG_074708.1, NG_074709.1, NG_076643.1, NG_077975.1, NG_077976.1, NG_080772.1, NG_080773.1, NG_081003.1, NG_081782.1, NG_088389.1, NG_148619.1, NG_157005.1, and NG_203390.1.

[0031] The fifth primer set can specifically detect a VIM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 218 to SEQ ID NO: 239. When the fifth primer set is a forward primer represented by SEQ ID NO: 17 and a reverse primer represented by SEQ ID NO: 18 (VIM primer set1), a VIM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 184 to SEQ ID NO: 224 is specifically detected, and when the fifth primer set is a forward primer represented by SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 20 (VIM primer set2), a VIM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 225 to SEQ ID NO: 239 is specifically detected.

[0032] Specifically, the fifth primer set specifically detects the VIM-2-like gene (NCBI GenBank No. NG_050347.1) or a variant thereof, and the variant of the VIM-2-like gene is selected from the group consisting of NCBI GenBank No. NG_062231.1, NG_157061.1, NG_065942.1, NG_065448.1, NG_050357.1, NG_050356.1, NG_076811.1, NG_079261.1, NG_050358.1, NG_050349.1, NG_050369.1, NG_050337.1, NG_050347.1, NG_050348.1, NG_064785.1, NG_070756.1, NG_050379.1, NG_050350.1, NG_061404.1, NG_050342.1, NG_074754.1, NG_070757.1, NG_050338.1, NG_050378.1, NG_078035.1, NG_050383.1, NG_050382.1, NG_064786.1, NG_050372.1, NG_203418.1, NG_050363.1, NG_088450.1, NG_203417.1, NG_050380.1, Any one selected from NG_050345.1, NG_050343.1, NG_050373.1, NG_063896.1, NG_050344.1, NG_050374.1, NG_064783.1, NG_060588.1, and NG_056405.1.

[0033] The sixth primer set can specifically detect an IMP gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 240 to SEQ ID NO: 278. When the sixth primer set is a forward primer represented by SEQ ID NO: 21 and a reverse primer represented by SEQ ID NO: 22 (IMP primer set1), an IMP gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 240 to SEQ ID NO: 269 is specifically detected, and when the sixth primer set is a forward primer represented by SEQ ID NO: 23 and a reverse primer represented by SEQ ID NO: 24 (IMP primer set2), an IMP gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 270 to SEQ ID NO: 278 is specifically detected.

[0034] Specifically, the above-mentioned 6th primer set specifically detects the IMP gene (NCBI GenBank No. NG_049172.1) or its mutation(variant), and the above-mentioned IMP gene mutation is NCBI GenBank No. NG_049172.1, NG_049173.1, NG_049189.1, NG_049190.1, NG_049194.1, NG_049195.1, NG_049199.1, NG_049202.1, NG_049203.1, NG_049204.1, NG_049206.1, NG_049214.1, NG_049217.1, NG_055477.1, NG_049220.1, NG_050945.1, NG_051166.1, NG_054676.1, NG_061409.1, NG_061410.1, NG_061411.1, NG_061626.1, NG_070737.1, NG_070738.1, NG_088392.1, NG_148620.1, NG_055665.1, NG_057607.1, NG_242291.1, NG_231544.1, NG_049192.1, NG_049207.1, NG_049212.1, NG_049213.1, NG_049221.1, NG_065947.1, NG_066696.1, NG_079229.1, NG_049178.1, NG_049193.1, NG_049209.1, NG_049215.1, NG_051513.1, NG_065873.1, NG_049223.1, NG_056414.1, NG_057463.1, NG_049174.1, NG_049179.1, NG_049185.1, NG_049186.1, NG_049205.1, NG_049208.1, NG_049219.1, NG_055584.1, NG_079228.1, NG_062274.1, NG_049176.1, NG_049180.1, NG_049182.1, NG_049183.1, NG_049184.1, NG_049187.1, NG_049188.1, NG_049191.1, NG_049198.1, NG_049201.1, NG_064725.1, NG_054710.1, NG_055271.1, NG_049222.1, NG_065875.1, NG_079887.1, NG_080776.1, NG_056176.1, NG_049177.1, NG_049181.1, NG_049197.1, NG_064724.1, NG_049210.1, NG_049211.1, NG_049216.1, NG_049218.1, NG_066508.1, NG_057606.1, NG_065874.1, NG_076650.1, NG_076651.1, NG_076634.1, NG_157006.1, NG_203391.1, NG_049175.1, NG_242292.1, NG_052049.1, NG_074713.1, NG_049196.1, Any one selected from NG_049200.1, and NG_079227.1.

[0035] The above KPC (Klebsiella pneumoniaecarbapenemase) is a β-lactamase carbapenemase belonging to class A, and is the most common carbapenemase worldwide, including in the United States and Europe. It is most commonly found in Klebsiella pneumoniae, but is also identified in most Enterobacterales, including Enterobacter spp. and Escherichia coli. In Korea, KPC-2 is the most common, and KPC-3 and KPC-4 are also frequently identified. As of March 2024, 194 variants exist.

[0036] The above-mentioned NDM (New Delhi metallo-beta-lactamase) is a class B metallo-beta-lactamase carbapenemase that hydrolyzes most beta-lactam antibiotics, including penicillins, cephalosporins, and carbapenems, except for aztreonam. It was first identified in Klebsiella pneumoniae and Escherichia coli in 2008, and although it appeared later than KPC, its rapid global spread has become a problem. As of March 2024, 60 variants exist.

[0037] The above OXA (oxacillinase)-48 is a β-lactamase carbapenemase of the OXA (oxacillinase) family belonging to class D, which has a high hydrolytic ability for penicillin, and hydrolyzes carbapenems unlike other OXA families. It has low genetic similarity with OXA-10, OXA-23, OXA-40, and OXA-1, and unlike most OXA families which are mainly found in Acinetobacter species, the acquisition of OXA-48 family genes is only found in Enterobacterales including Klebsiella pneumoniae. In Korea, OXA-181 and OXA-232 of the OXA-48 family are most commonly confirmed. As of March 2024, there are 1,182 OXA variants, of which 58 variants are OXA-48.

[0038] The GES gene, a class A β-lactamase carbapenemase, has been reported in Acinetobacter baumannii, Pseudomonas aeruginosa, and various Enterobacterales in South America, the Middle East, Southeast Asia, Africa, and Europe. Although its frequency among CPEs is relatively low in Korea, GES-5 continues to occur. As of March 2024, 59 variants exist.

[0039] The above VIM (Verona integron-encoded metallo-β-lactamase)-2 is a class B metallo-β-lactamase carbapenemase, first discovered in Pseudomonas aeruginosa. As of March 2024, there are 84 VIM variants, which are classified into VIM-1-like, VIM-2-like, and VIM-7-like according to genetic similarity. VIM is mainly common in Europe, and the incidence in Korea is very low. It is known that VIM-2Pseudomonas aeruginosa is widely distributed worldwide, and the occurrence of VIM-2-like series such as VIM-3 and VIM-6 is mainly found in Asia, including Korea. There are 43 VIM-2-like variants.

[0040] The IMP (imipenemase) gene above is a class B metallo-β-lactamase carbapenemase that inactivates most beta-lactam antibiotics except monobactams. Since IMP-1 was discovered from Pseudomonas aeruginosa in Japan, it has spread to enteric bacteria through plasmid-mediated transmission. As of March 2024, 98 variants have been identified, which are divided into at least 7 phylogenetic clusters (G1 to G7). IMP G1 contains the largest number of 28 variants as an IMP-1 / 4-like gene, followed by IMP G2 and G4 with 18 variants each. In addition, IMP G5 with 16 variants and G3 with 10 variants have been identified, while G6 and G7 contain 5 and 3 variants, respectively. In Korea, there are reports of the occurrence of IMP-1 and IMP-4 of G1 (IMP-1 / 4-like), and the incidence is somewhat low along with GES.

[0041] The above composition is for single PCR, triplex PCR, multiplex PCR, real time PCR, triplex real time PCR, or multiplex real time PCR.

[0042] In one embodiment of the present invention, the first primer set is composed of a forward primer represented by SEQ ID NO: 1 and a reverse primer represented by SEQ ID NO: 2, the second primer set is composed of a forward primer represented by SEQ ID NO: 5 and a reverse primer represented by SEQ ID NO: 6, the third primer set is composed of a forward primer represented by SEQ ID NO: 9 and a reverse primer represented by SEQ ID NO: 10, the fourth primer set is composed of a forward primer represented by SEQ ID NO: 13 and a reverse primer represented by SEQ ID NO: 16, the fifth primer set is composed of a forward primer represented by SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 20, and the sixth primer set is composed of a forward primer represented by SEQ ID NO: 21 and a reverse primer represented by SEQ ID NO: 22.

[0043] In one embodiment of the present invention, a composition for detecting carbapenemase producing Enterobacterales (CPE) comprises the first primer set, the second primer set, and the third primer set, wherein the first primer set is composed of a forward primer represented by SEQ ID NO: 1 and a reverse primer represented by SEQ ID NO: 2, the second primer set is composed of a forward primer represented by SEQ ID NO: 5 and a reverse primer represented by SEQ ID NO: 6, and the third primer set is composed of a forward primer represented by SEQ ID NO: 9 and a reverse primer represented by SEQ ID NO: 10, and the composition is for triplex PCR or triplex real time PCR. When the above composition is for triplex real time PCR, it additionally includes a first probe represented by SEQ ID NO: 27, a second probe represented by SEQ ID NO: 28, and a third probe represented by SEQ ID NO: 29.

[0044] In another embodiment of the present invention, a composition for detecting carbapenemase producing Enterobacterales (CPE) comprises the fourth primer set, the fifth primer set, and the sixth primer set, wherein the fourth primer set is composed of a forward primer represented by SEQ ID NO: 13 and a reverse primer represented by SEQ ID NO: 16, the fifth primer set is composed of a forward primer represented by SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 20, the sixth primer set is composed of a forward primer represented by SEQ ID NO: 21 and a reverse primer represented by SEQ ID NO: 22, and the composition is for triplex PCR or triplex real time PCR. When the above composition is for triplex real time PCR, it additionally includes a fourth probe represented by SEQ ID NO: 30, a fifth probe represented by SEQ ID NO: 31, and a sixth probe represented by SEQ ID NO: 32.

[0045] In the present invention, the probe has a fluorophore bound to the 5' end and a quencher bound to the 3' end.

[0046] The above fluorophores are fluorescein, 6-carboxyfluorescein (FAM, 6-carboxyfluorescein), hexachloro-6-carboxyfluorescein (HEX, hexachloro-6-carboxyfluorescein), tetrachloro-6-carboxyfluorescein (TET, tetrachloro-6-carboxyfluorescein), 2-chloro-7-phenyl-1,4-dichloro-6-carboxyfluorescein (VIC, 2-chloro-7-phenyl-1,4-dichloro-6-carboxyfluorescein), 2,7-dimethoxy-4,5-dichloro-6-carboxyfluorescein (JOE, 2,7-dimethoxy-4,5-dichloro-6-carboxyfluorescein), It may be any one selected from 5-((2-aminoethyl)amino)naphthalene-1-sulfonic acid, coumarin and coumarin derivatives, cyanine-5 (Cy5), lucifer yellow, Texas red, tetramethylrhodamine, Yakima Yellow (YG), and Cal Fluor Red 610 (CFR).

[0047] The above quencher may be any one selected from tetramethylrhodamine (TAMRA), 4-(4-dimethylaminophenylazo)benzoic acid, 4-dimethylaminophenylazophenyl-4-maleimide, carboxytetramethylrhodamine, Black Hole Quencher 1 (BHQ1), Black Hole Quencher 2 (BHQ2), and BHQ dyes.

[0048] In one embodiment of the present invention, when performing a triplex real-time PCR using the first primer set, the third primer set, and the fourth primer set, a first PCR amplification product is formed by the first primer set, a second PCR amplification product is formed by the second primer set, and a third PCR amplification product is formed by the third primer set. The first PCR amplification product has a size of 130 bp and specifically binds to the first probe represented by SEQ ID NO: 27, the second PCR amplification product has a size of 242 bp and specifically binds to the second probe represented by SEQ ID NO: 28, and the third PCR amplification product has a size of 111 bp and specifically binds to the third probe represented by SEQ ID NO: 29. The 5'-ends of the first probe, the second probe, and the third probe are bound to fluorophores, and in one embodiment of the present invention, when the first probe, the second probe, and the third probe are mixed together in one PCR cocktail, the fluorophores bound to the first probe, the second probe, and the third probe are not identical. More specifically, when triplex real-time PCR is performed using the first primer set, the third primer set, and the fourth primer set, 6-carboxyfluorescein (FAM) is bound to the 5'-end of the first probe mixed together, Texas red is bound to the 5'-end of the second probe, and cyanine-5 (Cy5) is bound to the 5'-end of the third probe.

[0049] In another embodiment of the present invention, when a triplex real-time PCR is performed using the fourth primer set, the fifth primer set, and the sixth primer set, a fourth PCR amplification product is formed by the fourth primer set, a fifth PCR amplification product is formed by the fifth primer set, and a sixth PCR amplification product is formed by the sixth primer set. The fourth PCR amplification product has a size of 267 bp and specifically binds to the fourth probe represented by SEQ ID NO: 30, the fifth PCR amplification product has a size of 281 bp and specifically binds to the fifth probe represented by SEQ ID NO: 31, and the sixth PCR amplification product has a size of 261 bp and specifically binds to the sixth probe represented by SEQ ID NO: 32. The 4th probe, the 5th probe, and the 6th probe are coupled with fluorophores at the 5' end, and in one embodiment of the present invention, when the 4th probe, the 5th probe, and the 6th probe are mixed together in one PCR cocktail, the fluorophores coupled to the 4th probe, the 5th probe, and the 6th probe are not identical. More specifically, when triplex real-time PCR is performed using the 4th primer set, the 5th primer set, and the 6th primer set, cyanine-5 (Cy5) is coupled to the 5' end of the 4th probe that is mixed together, 6-carboxyfluorescein (FAM) is coupled to the 5' end of the 5th probe, and Texas red is coupled to the 5' end of the 6th probe.

[0050] In addition, the present invention comprises a step of isolating DNA from a sample (step 1); a step of performing a polymerase chain reaction (PCR) using at least one primer set selected from the group consisting of a first primer set comprising a forward primer represented by SEQ ID NO: 1 and a reverse primer represented by SEQ ID NO: 2, using the DNA separated in the first step as a template, a second primer set comprising a forward primer represented by SEQ ID NO: 5 and a reverse primer represented by SEQ ID NO: 6, a third primer set comprising a forward primer represented by SEQ ID NO: 9 and a reverse primer represented by SEQ ID NO: 10, a fourth primer set comprising a forward primer represented by SEQ ID NO: 13 and a reverse primer represented by SEQ ID NO: 16, a fifth primer set comprising a forward primer represented by SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 20, and a sixth primer set comprising a forward primer represented by SEQ ID NO: 21 and a reverse primer represented by SEQ ID NO: 22; a step of performing electrophoresis on a PCR amplification product obtained in the second step (step 3); And a step (fourth step) of detecting a carbapenemase-producing Enterobacterales (CPE) strain according to the presence or absence of a PCR amplification product confirmed in the third step is provided. The present invention provides a method for detecting a carbapenemase-producing Enterobacterales (CPE) strain.

[0051] In the second step, the polymerase chain reaction (PCR) can be performed under conditions in which the annealing temperature is 50.0°C to 59.1°C, and when triplex PCR is performed in the second step, the annealing temperature can be performed under conditions in which the annealing temperature is 56°C to 57°C.

[0052] In the fourth step, the size of the amplification product (amplicon) obtained in the second step may be 111 bp to 281 bp, and if a band having a size of 130 bp is present in the PCR amplification product, it is determined that a CPE strain having a KPC gene or a variant thereof is present in the sample, if a band having a size of 111 bp is present in the PCR amplification product, it is determined that a CPE strain having an OXA-48 gene or a variant thereof is present in the sample, if a band having a size of 242 bp is present in the PCR amplification product, it is determined that a CPE strain having an NDM gene or a variant thereof is present in the sample, if a band having a size of 267 bp is present in the PCR amplification product, it is determined that a CPE strain having a GES gene or a variant thereof is present in the sample, and if a band having a size of 281 bp is present in the PCR amplification product, it is determined that a VIM-2 gene or a variant thereof is present in the sample. It is determined that a CPE strain having a variant exists, and if a band with a size of 261 bp exists in the PCR amplification product, it is determined that a CPE strain having an IMP gene or a variant thereof exists in the sample.

[0053] In one embodiment of the present invention, in the second step, a triplex PCR or a triplex real-time PCR may be performed using the first primer set, the second primer set, and the third primer set. When a triplex real-time PCR is performed in the second step, a first probe represented by SEQ ID NO: 27, a second probe represented by SEQ ID NO: 28, and a third probe represented by SEQ ID NO: 29 are additionally used.

[0054] In addition, in another embodiment of the present invention, in the second step, a triplex PCR or a triplex real time PCR may be performed using the fourth primer set, the fifth primer set, and the sixth primer set, and when a triplex real time PCR is performed in the second step, a fourth probe represented by SEQ ID NO: 30, a fifth probe represented by SEQ ID NO: 31, and a sixth probe represented by SEQ ID NO: 32 are additionally used.

[0055] In all embodiments of the present invention, the method may further include a seventh primer set for amplifying 16S rRNA to determine the presence or absence of a strain in a sample, wherein the seventh primer set comprises a forward primer represented by SEQ ID NO: 25 and a reverse primer represented by SEQ ID NO: 26. In addition, when performing real-time polymerase chain reaction (real-time PCR) including the seventh primer set, the method may further include a seventh probe represented by SEQ ID NO: 33, wherein the seventh probe has cyanine-3 (Cy3) bound to its 5' end.

[0056] In the present invention, the "primer" refers to an oligonucleotide, which can act as an initiator of synthesis under conditions that induce the synthesis of a primer extension product complementary to a nucleic acid chain (template), i.e., the presence of nucleotides and a polymerization agent such as DNA polymerase, and conditions of suitable temperature and pH. Preferably, the primer is a deoxyribonucleotide and is single-stranded. The primer used in the present invention may include naturally occurring dNMPs (i.e., dAMP, dGMP, dCMP, and dTMP), modified nucleotides, or non-natural nucleotides. In addition, the primer may also include ribonucleotides. The primer must be sufficiently long to prime the synthesis of an extension product in the presence of a polymerization agent. The appropriate length of the primer is determined by several factors, such as temperature, application, and the source of the primer, but is typically 15-30 nucleotides. Shorter primer molecules generally require lower temperatures to form sufficiently stable hybrid complexes with the template. The terms annealing or priming refer to the juxtaposition of an oligodeoxynucleotide or nucleic acid to a template nucleic acid, which allows a polymerase to polymerize the nucleotides to form a nucleic acid molecule complementary to the template nucleic acid or a portion thereof.

[0057] The "primer set" of the present invention refers to a pair of primer sets consisting of a forward primer and a reverse primer, and is a primer set that amplifies all or part of a specific gene by a conventional PCR method. The primer set amplifies all or part of one gene per set, and when two or more of the primer sets are included, it can be used for the purpose of amplifying all or part of two or more target genes. When two or more of the primer sets are used, they are designed in a combination that does not form a primer dimer, and are designed to amplify only all or part of the target gene and not form other amplification products, and are designed so that the sizes of the amplification products amplified by each primer set are different, so that they can be distinguished on electrophoresis.

[0058] In the present invention, a "probe" refers to a nucleic acid fragment, such as DNA, ranging from a few bases to several hundred bases, that can specifically bind to DNA and is labeled to enable detection of the presence or absence of a specific DNA and its expression level. The probe can be produced in the form of an oligonucleotide probe, a single-stranded DNA probe, a double-stranded DNA probe, an RNA probe, etc. The selection of an appropriate probe and hybridization conditions can be appropriately selected according to techniques known in the art.

[0059] The concentration of the above primer or probe can be selected and used in various ways by a person skilled in the art depending on the experimental conditions, and is preferably 1 to 1000 nM each, and more preferably 100 to 500 nM each, but is not limited thereto.

[0060] The step of performing the above real-time polymerase chain reaction may be performed under conditions of 30 to 50 cycles, 30 to 46 cycles, 30 to 44 cycles, 33 to 50 cycles, 33 to 46 cycles, 33 to 44 cycles, 36 to 50 cycles, 36 to 46 cycles, 36 to 44 cycles, 38 to 50 cycles or 38 to 46 cycles, for example, 38 to 44 cycles, but is not limited thereto.

[0061] Hereinafter, to aid understanding of the present invention, examples will be given in detail. However, the following examples are intended only to illustrate the scope of the present invention and are not intended to limit its scope. These examples are provided to more fully explain the present invention to those of average skill in the art.

[0062]

[0063] Example 1. Primer design and in silico testing

[0064] We designed a set of genetic primers that can broadly detect variants of carbapenemase-producing Enterobacterales (CPE). As of March 2024, the sequences of 194 KPC, 60 NDM, 58 OXA-48-family (1182 OXAs), 59 GES, 43 VIM-2-like (84 VIMs), and 28 IMP G1 (98 IMPs) genes related to CPE registered in the Pathogen Detection Reference Gene Catalog of the National Center for Biotechnology Information (NCBI) were downloaded and compared using ClustalW Multi alignment (Booststrap NJ Tree 1000).

[0065] A primer set capable of broadly detecting KPC, GES, NDM, OXA-48 family, VIM-2-like, and IMP G1 (IMP-1 / 4-like) genes targeting regions with high homology in the aligned gene sequence was designed as shown in Table 1 below. The primers were designed with a GC ratio of 40.9% to 55.0%, a Tm (Melting Temperature) of 50.0℃ to 55.1℃, and amplicon sizes of 111 bp to 281 bp. Two forward primers and two reverse primers were designed for each gene. Primers f_1 and r_1 were used as primer set1, and f_2 and r_2 were combined to form primer set2. f_1 and r_2, and f_2 and r_1 were combined to form primer mix1 and primer mix2.

[0066] Target genesPrimersSequences (5'->3')GC(%)Tm(℃)AmpliconSize (bp)Set*NameKPCSet1kpc_f_1 (SEQ. 01)RTTCTGCTGTCTTGTYTYTC50.050.2130kpc_r_1 (SEQ. 02)TATCYATCGCGTACACACC52.651.4Set2kpc_f_2 (SEQ. 03)ATACAGTGATAACGCCGCC52.652264kpc_r_2 (SEQ. 04)GTCGTGTTTCCCTTTAGCC52.650.6NDMSet1ndm_f_1 (SEQ. 05)TTGCBCAATATTATGCACCC45.050.3242ndm_r_1 (SEQ. 06)CCATCCCTGACGATCAAAC52.650.0Set2ndm_f_2 (SEQ. 07)TGGATCAAGCAGGAGATCAAC47.651.9126ndm_r_2 (SEQ. 08)CGACAACGCATTGGCATAAG50.052.6OXA-48Set1oxa48_f_1 (SEQ. 09)TGGAATGAGAATAAGCAGCAAG40.950.9111oxa48_r_1 (SEQ. 10)CAAATCGAGGGCGATCAAG52.650.9Set2oxa48_f_2 (SEQ. 11)TTCGGCYAYBSAGCAAATC52.652.1190oxa48_r_2 (SEQ. 12)YTCAACCCAACCRACCCAC52.652.3GESSet1ges_f_1 (SEQ. 13)CTGCAATGACGCAGTATTTTC42.950.6266ges_r_1 (SEQ. 14)TTTCTCTCCAACAACCCAATC42.950.2Set2ges_f_2 (SEQ. 15)CGTACTGTGGCHAAARTCCTC52.452.4142ges_r_2 (SEQ. 16)TTTTCTCTCCAACAACCCAATC40.950.9VIM-2Set1vim2_f_1 (SEQ. 17)CGACAGTCAGCGAAATTCC52.651.2150vim2_r_1 (SEQ. 18)CAATCAAAAGCAACTCATCACC40.950.8Set2vim2_f_2 (SEQ.19)TGCAGTCTCCACGCACTTTC55.055.1281vim2_r_2 (SEQ. 20)ACTCATAAATCGCACAACCACC45.553.4IMP G1Set1imp1_f_1 (SEQ. 21)GCTTAWTTCTCRATCYATCCCC45.551.0261imp1_r_1 (SEQ. 22)GCTTCTAWATTTGCGTCACY45.050.1Set2imp1_f_2 (SEQ. 23)TCATTTTCATAGYGACAGCAC42.951.3227imp1_r_2 (SEQ. 24)TCCTTTYAGGCARCCAAAC47.450.2*Set1: f_1&r_1, set2: f_2&r_2, mix1: f_1&r_2, mix2: f_2&r_1.

[0067]

[0068] To determine whether the designed primer set above is capable of PCR, validation was performed on the in silico simulation site (insilico.ehu.eus). As shown in Table 2 below, the in silico validation results confirmed that the primer set could detect all CPE variants confirmed to occur in Korea. For KPC, NDM, VIM-2-like, and IMP G1, all variants could be detected, and for GES, all variants could be detected except for GES-23, which had a very low detection frequency. Primer set 1 of the OXA-48 family could detect 54 variants, including OXA-48 and OXA-181, except for OXA-54, OXA-436, OXA-535, and OXA-833, and primer set 2 could detect 57 variants except for OXA-436. The VIM-2-like gene was able to detect 43 variants, and no variants were detected in the VIM-1-like and VIM-7-like genes. In the case of IMP G1, primer set 1 detected a total of 43 variants in the G2~G7 groups other than the G1 group, including 17 in G2, 14 in G4, 8 in G5, 1 in G6, and 3 in G7. In the case of IMP G1 primer set 2, no variants in the G2~G7 groups other than the G1 group were detected.

[0069] Target geneVariants (N)PrimerIn silico testundetectable variants1bp mismatch2bp mismatchKPC194Set10(f) KPC-147-Set20(f) KPC-109, 179(r) KPC-131, 149-GES59Set1,2Mix1,21 (GES-23)(r) GES-32, 41, 44, 59-NDM60Set1,20--OXA-48 family58Set14 (OXA-54, 436, 535, 833)--1 (OXA-436)-(f) OXA-54(r) OXA-535, 833VIM-2-like43Set10(f)VIM-30, 45, 46, 65, 67Set20(r)VIM-31IMP G127Set10Set20

[0070]

[0071] Example 2. Establishment of PCR conditions

[0072] In order to establish the PCR conditions of the primer set produced in the above Example 1, gradient PCR (annealing temperature: 50℃ to 59.1℃) was performed for each gene. Other than the conditions for each PCR step, general PCR was performed under the conditions shown in Table 3 below, and real-time PCR was performed under the conditions shown in Table 4 below. The template was used by extracting the nucleic acid of the strain in which the target gene was confirmed with InstaGene Matrix, and PCR was performed using AccuPower® PCR PreMix according to the manual. All primers were used at a concentration of 10 pmole / rxn.

[0073] PCR StepTemperatureTimeCyclesPre-Denaturation95℃5 min1 cycleDenaturation95℃20 sec35 cycleAnnealing50~59.1℃20 secExtension72℃1 minFinal Extension72℃5 min1 cycleEnd4℃-

[0074] Real-time PCR StepTemperatureTimeCyclesPre-Denaturation95℃3 min1 cycleDenaturation95℃5 sec35 cycleAnnealing50~59.1℃30 secEnd4℃-

[0075] As a result of confirming the band of the PCR product through 1.5% agarose gel electrophoresis, the reaction was strong at an annealing temperature of 51.9℃ to 56.1℃ under both general PCR and real-time PCR conditions, and the nonspecific band decreased as the temperature increased (Fig. 1). Based on the two results, one primer set with good reactivity among the four primer sets was selected and used for multiplex PCR. KPC, NDM, OXA-48, and IMP G1 showed the best results with primer set 1, VIM-2 showed the best results with primer set 2, and GES showed the best results with primer mix 1.

[0076]

[0077] Example 3. Establishment of Triplex PCR Conditions

[0078] In order to perform multiplex PCR to detect three genes at once by combining the primer sets selected in Example 2, two triplex PCR sets were configured as shown in Table 5 below. The primer sets of KPC, NDM, and OXA-48, which are frequently isolated in Korea, were configured as triplex PCR set 1, and six primers of GES, VIM-2, and IMP G1, which are relatively frequently isolated, were configured as triplex PCR set 2. 16S rRNA primer was used as an internal control for each set. The template was used by extracting the nucleic acid of the strain in which the target gene was confirmed with InstaGene® Matrix, and PCR was performed using AccuPower® PCR PreMix according to the manual. All primers were used at a concentration of 5.0 pmole / rxn. The triplex PCR conditions were as shown in Table 6.

[0079] Triplex PCR setTarget genesPrimers (5'->3')Amplicon size(bp)Triplex PCR set1KPCkpc_f_1 (SEQ. 01) & kpc_r_1 (SEQ. 02)130NDMndm_f_1 (SEQ. 05) & ndm_r_1 (SEQ. 06)242OXA-48oxa48_f_1 (SEQ. 09) & oxa_48_r_1 (SEQ. 10)111Triplex PCR set2GESges_f_1 (SEQ. 13) & ges_r_2 (SEQ. 16)267VIM-2vim2_f_2 (SEQ. 19) & vim2_r_2 (SEQ. 20)281IMP G1imp1_f_1 (SEQ. 21) & imp1_r_1 (SEQ. 22)261Internal control16S rRNA16s_f (SEQ. 25)(TCCTACGGGAGGCAGCAGT)46616s_r (SEQ. 26)(GGACTACCAGGGTATCTAATCCTGTT)

[0080] StepTemperatureTimeCyclesPre-Denaturation95℃3 min1 cycleStep 1Denaturation95℃5 sec3 cycleAnnealing57℃50 secStep 2Denaturation95℃5 sec45 cycleAnnealing56℃30 secEnd4℃-

[0081] Two triplex PCR sets were verified for 11 strains (2 KPC, 2 GES, 3 NDM, 2 OXA-48, 1 VIM-2, 1 IMP-4) whose target genes and variant types were confirmed through sequencing and 1 strain without a carbapenemase gene. As a result, as shown in Fig. 2, bands corresponding to KPC, NDM, and OXA-48 in size were confirmed in triplex PCR set1, and bands corresponding to GES, VIM-2, and IMP in size were confirmed in triplex PCR set2.

[0082]

[0083] Example 4. Probe design and confirmation of optimal concentration for real-time PCR.

[0084] In order to apply the primer sets of each gene designed in Example 2 to real-time PCR, probes capable of detecting each gene were produced as shown in Table 7 below. A dye for fluorescent color development was attached to the 5' end of the sequence, and a quencher was attached to the 3' end of the sequence. The length of the produced probes was 23 to 39 mer, the GC ratio was 44.0% to 65.2%, and the annealing temperature was 57.8°C to 68.0°C, which differed from the primer set by a minimum of 6.8°C and a maximum of 14.8°C. The probes and primer sets designed for each gene were combined, and real-time PCR was performed under the conditions shown in Table 8 below according to the manual of TAKARA Probe qPCR Mix, with UNG. The template was used by extracting the nucleic acid of the strain in which the target gene was confirmed using InstaGene™ Matrix, and was used at a concentration of 1.0 ng / ul per reaction and 10 -5 Six serial dilutions were used up to 10 μg / mL. The probe was used at concentrations of 0.05, 0.1, 0.5, 1.0, 2.0, and 3.0 pmol / mL, respectively, and all primers were used at a concentration of 10.0 pmole / mL. Through this, the optimal probe concentration was determined, efficiency was confirmed, and the detection limit according to template concentration was measured.

[0085] ProbesSequences (5'Dye->3'Quencher)Length(mer)GC(%)Tm(℃)kpc_p (SEQ. 27)(FAM) TTTCTGCCACCGCGCTGACCAACCT (BHQ1)2550.064.9ndm_p (SEQ. 28)(Texas Red) MTCKCACCGAATGTCTGGCAGCACACTT (BHQ2)2857.164.9oxa48_p (SEQ. 29)(Cyanone5) AATCTTAAACGGGCGAACCAAGCAT (BHQ2)2544.057.8ges_p (SEQ. 30)(Cyanine5) CGACAACACACCTGGCGACCTCAGARATACAA (BHQ2)3253.164.6vim2_p (SEQ. 31)(FAM) CGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTG (BHQ1)3953.868.0imp1_p (SEQ. 32)(Texas Red) TTTATCCAGGCCCRGGACACACTCCA (BHQ2)2653.861.116s_p (SEQ. 33)(Cyanine3) CGTATTACCGCGGCTGCTGGCAC (BHQ1)2365.267.6

[0086] StepTemperatureTimeCyclesPre-Denaturation95℃3 min1 cycleStep 1Denaturation95℃5 sec3 cycleAnnealing57℃50 secStep 2Denaturation95℃5 sec45 cycleAnnealing56℃30 secEnd4℃-

[0087] As shown in Fig. 3, real-time PCR results confirmed the detection of the target gene in all reactions except for GES reacted with a probe concentration of 0.05 pmol / rxn. As the probe concentration increased, the ΔRn value at which the reaction was detected increased. For 0.05 and 0.10 pmol / rxn, ΔRn was very low, making it difficult to set a threshold baseline. At concentrations of 0.50 pmol / rxn or higher, ΔRn of all genes was confirmed to be over 50,000.

[0088] When calculating the efficiency of each gene according to the probe concentration, as shown in Table 9 below, Gating and NDM were confirmed to be lower than other genes, and the detection of IMP was confirmed to be high overall regardless of the probe concentration. The efficiency of the probe according to the template concentration was confirmed to be 0.88 to 0.94 on average. Excluding 0.05 pmol / rxn where GES was not detected, the efficiency was high overall over the range of 0.10 to 3.00 pmol / rxn, and the efficiency was highest at 0.50 and 2.00 pmol / rxn.

[0089] Target gene probe (pmol / rxn)0.050.100.501.002.003.00KPC0.930.990.970.810.920.92NDM0.660.690.670.610.620.5 8OXA-481.060.931.031.090.991.01GES-0.740.880.921.000.98VIM-20.940.920.940.880.980.87IMP G11.111.001.031.021.041.06Average0.940.880.920.890.920.90

[0090] *Efficiency=(10^(-1 / slope))-1, Slope=log(Template concentration) ΔCt graph slope

[0091] Example 5. Establishment of Triplex real-time PCR conditions

[0092] To establish a multiplex real-time PCR method capable of detecting six CPE genes, two triplex real-time PCR sets were constructed. Triplex real-time PCR set 1 was created by combining triplex PCR set 1 with KPC, OXA-48-family, and NDM probes, and triplex real-time PCR set 2 was created by combining triplex PCR set 2 with GES, VIM-2-like, and IMP G1 probes (Table 10). 16S rRNA primers and probes were used as internal controls for each set. Primers were used at a concentration of 5.0 pmole / rxn, and probes were used at a concentration of 1.0 pmole / rxn. The template was extracted from the nucleic acid of the strain in which the target gene was confirmed using InstaGene Matrix, and real-time PCR was performed under the conditions shown in Table 11 below according to the manual of TAKARA Probe qPCR Mix, with UNG. To reduce ΔCt and increase detection efficiency of real-time PCR, the number of cycles in step 1 was adjusted to 3, 8, and 10.

[0093] Triplex real-time PCR setTarget genePrimersProbesAmplicon size(bp)Triplex real-time PCR set1KPCkpc_f_1 & kpc_r_1kpc_p130NDMndm_f_1 & ndm_r_1ndm_p242OXA-48oxa48_f_1 & oxa_48_r_1oxa48_p111Triplex real-time PCR set2GESges_f_1 & ges_r_2ges_p267VIM-2vim2_f_2 & vim2_r_2vim2_p281IMP G1imp1_f_1 & imp1_r_1imp1_p261Internal control16S rRNA16s_f & 16s_r16s_p466

[0094] StepTemperatureTimeCyclesPre-Denaturation95℃3 min1 cycleStep 1Denaturation95℃5 sec3, 8, 10 cycleAnnealing57℃50 secStep 2Denaturation95℃5 sec45 cycleAnnealing56℃30 secEnd4℃-

[0095] Triplex real-time PCR set1 and Triplex real-time PCR set2 detected only the target genes, and no false positives or false negatives were observed. The average ΔCt according to the cycle number of Step 1 was 15.8 (13.4-17.5), 11.2 (9.2-14.4), and 9.3 (7.1-12.6) at 3, 8, and 10 cycles, respectively, confirming that the detection efficiency increased as the cycle number of Step 1 increased (Table 12). However, as the cycle number of Step 1 increased, the ΔRn of VIM tended to decrease rapidly (Fig. 4). Therefore, in this study, considering the ΔRn value of VIM, it was confirmed that the cycle number of Step 1 was appropriate to be 3-8.

[0096] Target geneProbeTriplex real-time PCR step1-cycles (ΔCt)3 cycles8 cycles10 cyclesKPC-2KPC15.59.87.4NDM-1NDM17.514.412.6OXA-181OXA-4815.312.510.3GES- 5GES17.110.38.3VIM-2VIM-213.49.27.2IMP-4IMP-115.710.77.1Average15.811.29.3

[0097]

[0098] Example 6. Measurement of the detection limit of Triplex real-time PCR

[0099] The detection limit of the triplex real-time PCR, which established the optimal conditions, was confirmed. Experiments were conducted on six clinical isolates containing the target gene, and nucleic acids were extracted using InstaGene Matrix. The gene copy number of the extracted strains was calculated using the average gene length of the strains registered in NCBI, and experiments were conducted at a total of six concentration levels from 1.0X10^6 copies / ul to 1.0 copies / ul (Table 13). The composition of each triplex real-time PCR is as shown in Table 10, and the primers were used at a concentration of 5.0 pmole / rxn and the probe was used at a concentration of 1.0 pmole / rxn. The triplex real-time PCR conditions are as shown in Table 11, and the cycles of step 1 were 3 and 8, respectively.

[0100] SpeciesTarget geneGenome length (bp)DNA Con. (ng / ul)Klebsiella pneumoniaeKPC-25596280143.9Citrobacter freundiiNDM-15258000139.7Enterobacterspp.OXA-484811860239.8Enterobacterspp.GES-54811860243.5Enterobacterspp.IMP-44811860287.7Klebsiella pneumoniaeVIM-25596280159.8

[0101] When the cut-off value of ΔCt was set to 32, under the step1 3-cycle condition, IMP-4 could be detected up to at least 1 copy, VIM-2, KPC-2, OXA-181, and GES-5 could be detected up to at least 10 copies, and NDM-1 could be detected up to at least 100 copies (Table 14, Fig. 5). On the other hand, under the step1 8-cycle condition, all genes could be detected up to at least 1 copy except for NDM-1, which could be detected up to at least 10 copies. The average efficiency also showed an increase in the step1 8-cycle condition (0.97) compared to the step1 3-cycle condition (0.92). The average ΔCt value confirmed in each copy was lower by more than ΔCt 8.0 in the step1 8-cycle condition compared to the step1 3-cycle condition. In the Triplex real-time PCR amplification plot, the ΔRn values ​​of low gene copy numbers also showed a pattern of greatly increasing in all genes (Fig. 5).

[0102] Triplex real-time PCRTarget geneGene copy number (ΔCt)Efficiency110^110^210^310^410^5Step1_3cycleKPC-234.830.527.221.820.216.70.89NDM-136.934.930.726.222.217.60.79OXA-18133.930.82 8.224.521.316.90.99GES-534.530.126.923.319.916.40.90VIM-233.429.5 25.322.318.815.20.90IMP-429.526.723.320.117.013.51.04Average33.830 .426.923.019.916.10.92Step1_8cycleKPC-223.320.316.913.110.26.81.0 0NDM-133.929.023.919.214.19.80.61OXA-18126.123.019.516.012.69.30.9 7GES-524.321.017.414.511.48.01.04VIM-223.520.717.113.510.06.71.23 IMP-423.620.216.713.110.06.70.97Average25.822.418.614.911.47.90.97

[0103]

[0104] Example 7. Validation of triplex real-time PCR using clinical isolates

[0105] The performance of the triplex real-time PCR of Example 3 was verified for 132 strains of 9 strains confirmed to have CPE according to the Kor-GLASS manual (Table 15) among the CRE strains isolated at Inje University Busan Paik Hospital. 47 strains harboring KPC-2 and 3, 49 strains harboring NDM-1, 5 and 7, 15 strains harboring OXA-48 and 181, 9 strains harboring GES-5 and 6, 1 strain harboring VIM-2, 1 strain harboring IMP-4, 3 strains harboring 2-3 genes simultaneously, and 58 negative control strains without CPE genes were included in the experiment (Table 16). The template was used by extracting the nucleic acid of each strain with InstaGene Matrix. The composition of each triplex real-time PCR is as shown in Table 10, and the primers were used at a concentration of 5.0 pmole / rxn and the probe was used at a concentration of 1.0 pmole / rxn. The triplex real-time PCR conditions are as shown in Table 11, and the cycle of step 1 was 3. The positive concordance, negative concordance, and overall concordance between the results derived through conventional PCR and sequencing and the triplex real-time PCR results were calculated as shown in Table 17. In case of discrepant results, the conventional PCR and sequencing were retested, and the CPE gene was reconfirmed through triplex real-time PCR retest and additional sequencing to derive the final result for each strain. Based on the final results, the sensitivity and specificity of the conventional PCR and sequencing test method and the developed triplex real-time PCR test method were calculated as shown in Table 18.

[0106] Target genesPrimer nameSequences (5'->3')Size (bp)KPCKPC F (SEQ. 34)ATGTCACTGTATCGCCGTCT893KPC R (SEQ. 35)TTTTCAGAGCCTTACTGCCCGESGES F (SEQ. 36)GCGCTTCATTCACGCACTAT753GES R (SEQ. 37)GCGTAATCTCTCTCCTGGGCNDMNDM-1 F (SEQ. 38)CAATATTATGCACCCGGTCG726NDM-1 R (SEQ. 39)ATCATGCTGGCCTTGGGGAAIMP R (SEQ. 41)TTAGTTGCTTGGTTTTGATGVIMVIM F (SEQ. 42)TGGTCTACATGACCGCGTCT766VIM R (SEQ. 43)CGACTGAGCGATTTGTGTGOXA-48OXA-48 F (SEQ. 44)TTGGTGGCATCGATTATCGG744OXA-48 R (SEQ. 45)GAGCACTTCTTTTGTGATGGC* 94℃ 5min + 30X (94℃ 30 sec + 56℃ 20 sec + 72℃ 30 sec) + 72℃ 7 min

[0107]

[0108] Single PCR & sequencingCFECESKOKPKVPRROSMSub-totalTotalKPC-259101111 23947KPC-3 11 5 1 8NDM-1105128 1 2749NDM-5 10 9 2 21NDM-7 1 1OXA-48 21 9 1215OXA-181 3 3GES-5 4 4 89GES-6 1 1VIM-2 1 11IMP-4 1 11NDM-5,OXA-181 2 23KPC-2, NDM-5, OXA-181 1 1Negative101211515 55858Total2545298601177183* CF, Citrobacter freundii; EC, Escherichia coli; ES, Enterobacter spp.; KO, Klebsiella oxytoca; KP, Klebsiella pneumoniae; KV, Klebsiella variicola; PR, Providencia rettgeri; RO, Raoultella ornithinolytica; SM, Serratia marcescens

[0109] Single PCR & sequencing Positive Negative Triplex real-time PCR Positive AB Negative CD * Positive agreement (%) = A / (A + C) 100 ** Negative agreement (%) = D / (B + D) 100 *** Overall agreement (%) = (A + D) / (A + B + C + D) 100

[0110] Triplex real-time PCR or Single PCR & sequencing Positive Negative Final result Positive AB Negative CD * Sensitivity (%) = A / (A+B) Х100 ** Specificity (%) = D / (C+D) Х100

[0111]

[0112] As a result of confirming the CPE genes of a total of 183 CRE strains, as shown in Table 19 below, it was possible to determine the presence of 1 to 3 CPE genes in more than 9 different strains through the developed triplex real-time PCR test method.

[0113] Single PCR & sequencingTriplex real-time PCRCFECESKOKPKVPRROSMSub- totalTotalKPC-2KPC59101111 23947KPC-3KPC 11 5 1 8NDM-1NDM105127 1 2649KPC, NDM* 1 1NDM-5NDM 10 9 1 20KPC, NDM, IMP* 1 1NDM-7NDM 1 1OXA-48OXA 21 9 1215OXA-181OXA 2 2NDM, OXA* 1 1GES-5GES 3 4 79KPC, GES* 1 1GES-6GES 1 1VIM-2VIM 1 11IMP-4IMP 1 11NDM-5, OXA-181NDM, OXA 2 23KPC-2, NDM-5, OXA-181KPC, NDM, OXA 1 1NegativeNega9101159 54958GES, IMP*1 1NDM* 1 1KPC* 1 6 7Total2545298601177183

[0114] In addition, as shown in Table 20 below, the overall concordance rate between the existing single PCR & sequencing test results and the triplex real-time PCR results was 95.1%, the positive concordance rate was 100%, and the negative concordance rate was 84.5%. The reason for the low negative concordance rate was that 9 cases were confirmed to be negative in the existing single PCR & sequencing test, but were positive in the developed triplex real-time PCR results. In these cases, the CPE gene was not detected in the existing single PCR & sequencing retest, but the CPE gene was confirmed as a result of performing single PCR and sequencing using the primers developed in this study. In addition, additional CPE genes were confirmed in 4 strains that were previously confirmed to have a single CPE gene.

[0115] Single PCR & sequencing Positive Negative Triplex real-time PCR Positive 1259 Negative 049 * Positive agreement (%) = A / (A+C) 100= 100.0 ** Negative agreement (%) = D / (B+D) 100= 84.5 *** Overall agreement (%) = (A+D) / (A+B+C+D) 100= 95.1

[0116] The final results of the CPE gene test for each strain were established through re-examination and sequencing for discrepant results, and the sensitivity and specificity of the existing single PCR & sequencing test method and the developed triplex real-time PCR test method for the final results were calculated, as shown in Table 21 below. Specificity was confirmed to be 100% for both experimental methods for 183 target strains, and sensitivity was confirmed to be 100% for the developed triplex real-time PCR test method, which was higher than the sensitivity of 93.3% for the existing single PCR & sequencing test method.

[0117] Single PCR & sequencingTriplex real-time PCRPositiveNegativePositiveNegativeFinal resultPositive12591340Negative049049Sensitivity (%)93.3100Specificity (%)100100

[0118]

[0119] Additional information

[0120] Table 22 below shows the variants of the KPC gene and their NCBI registration numbers.

[0121] Variant typeNCBI No.Variant typeNCBI No.Variant typeNCBI No.KPC-10NG_049243.1KPC-79NG_071205.1KPC-140NG_088400.1KPC-11NG_049244.1KPC-62NG_073465.1KPC-141NG_088401.1KPC-12NG_049245.1KPC-63NG_073466.1KPC-142NG_088402.1KPC-13NG_049246.1KPC-64NG_073467.1KPC-143NG_088403.1KPC-14NG_049247.1KPC-65NG_073468.1KPC-144NG_088404.1KPC-15NG_049248.1KPC-80NG_073469.1KPC-99NG_088405.1KPC-16NG_049249.1KPC-81NG_073470.1KPC-145NG_148622.1KPC-17NG_049250.1KPC-82NG_073471.1KPC-146NG_148623.1KPC-18NG_049251.1KPC-47NG_074714.1KPC-147NG_148624.1KPC-19NG_049252.1KPC-48NG_074715.1KPC-148NG_148625.1KPC-2NG_049253.1KPC-67NG_074716.1KPC-151NG_148626.1KPC-21NG_049254.1KPC-68NG_074717.1KPC-153NG_148627.1KPC-22NG_049255.1KPC-69NG_074718.1KPC-109NG_149659.1KPC-24NG_049256.1KPC-70NG_074719.1KPC-155NG_149660.1KPC-3NG_049257.1KPC-84NG_074720.1KPC-156NG_149661.1KPC-4NG_049258.1KPC-85NG_074721.1KPC-157NG_149662.1KPC-5NG_049259.1KPC-86NG_074722.1KPC-136NG_157007.1KPC-6NG_049260.1KPC-87NG_074723.1KPC-159NG_157008.1KPC-7NG_049261.1KPC-88NG_074724.1KPC-160NG_157009.1KPC-8NG_049262.1KPC-90NG_076666.1KPC-161NG_157010.1KPC-25NG_051167.1KPC-91NG_076667.1KPC-162NG_157011.1KPC-26NG_051469.1KPC-94NG_076680.1KPC-163NG_157012.1KPC-28NG_052581.1KPC-95NG_076681.1KPC-164NG_157013.1KPC-27NG_052862.1KPC-98NG_078032.1KPC-165NG_157014.1KPC-30NG_054685.1KPC-96NG_078037.1KPC-166NG_157015.1KPC-31NG_055494.1KPC-97NG_078038.1KPC-167NG_157016.1KPC-32NG_055495.1KPC-103NG_078051.1KPC-124NG_203393.1KPC-29NG_055580.1KPC-104NG_078052.1KPC-129NG_203394.1KPC-33NG_056170.1KPC-107NG_078053.1KPC-178NG_203395.1KPC-34NG_057447.1KPC-105NG_078054.1KPC-179NG_203396.1KPC-35NG_060524.1KPC-108NG_078055.1KPC-180NG_203397.1KPC-23NG_060569.1KPC-106NG_078056.1KPC-158NG_228670.1KPC-36NG_061389.1KPC-102NG_078063.1KPC-181NG_228671.1KPC-37NG_061612.1KPC-112NG_079230.1KPC-182NG_228672.1KPC-38NG_062357.1KPC-83NG_079231.1KPC-183NG_228673.1KPC-39NG_063841.1KPC-89NG_079232.1KPC-184NG_228674.1KPC-40NG_064726.1KPC-92NG_079233.1KPC-154NG_231545.1KPC-42NG_064727.1KPC-113NG_079888.1KPC-170NG_231546.1KPC-43NG_064728.1KPC-114NG_079889.1KPC-185NG_231547.1KPC-44NG_065427.1KPC-115NG_079890.1KPC-186NG_231548.1KPC-41NG_065876.1KPC-116NG_079891.1KPC-187NG_231549.1KPC-45NG_065877.1KPC-117NG_079892.1KPC-189NG_231550.1KPC-46NG_065878.1KPC-118NG_079893.1KPC-190NG_231551.1KPC-51NG_067224.1KPC-119NG_079894.1KPC-191NG_231552.1KPC-52NG_067225.1KPC-120NG_079895.1KPC-192NG_231553.1KPC-54NG_067226.1KPC-121NG_079896.1KPC-137NG_242182.1KPC-56NG_068016.1KPC-122NG_079897.1KPC-168NG_242183.1KPC-53NG_068176.1KPC-123NG_079898.1KPC-176NG_242184.1KPC-55NG_068177.1KPC-125NG_080778.1KPC-193NG_242185.1KPC-50NG_068507.1KPC-126NG_080779.1KPC-194NG_242186.1KPC-57NG_068508.1KPC-93NG_080780.1KPC-195NG_242187.1KPC-58NG_070177.1KPC-100NG_081070.1KPC-196NG_242188.1KPC-59NG_070178.1KPC-127NG_081071.1KPC-197NG_242189.1KPC-60NG_070179.1KPC-128NG_081072.1KPC-201NG_242190.1KPC-61NG_070180.1KPC-130NG_081699.1KPC-149NG_242293.1KPC-66NG_070739.1KPC-131NG_081700.1KPC-150NG_242294.1KPC-72NG_070740.1KPC-132NG_081783.1KPC-152NG_242295.1KPC-73NG_070741.1KPC-133NG_081784.1KPC-202NG_24 2296.1KPC-74NG_070742.1KPC-111NG_081791.1KPC-203NG_242297.1KPC-75NG_070743.1KPC-101NG_088394.1KPC-204NG_242298.1KPC-71NG _070895.1KPC-110NG_088395.1KPC-205NG_242299.1KPC-76NG_070896.1KPC-134NG_088396.1KPC-206NG_242300.1KPC-77NG_070897.1KPC- 135NG_088397.1KPC-207NG_242301.1KPC-49NG_071203.1KPC-138NG_088398.1KPC-208NG_242302.1KPC-78NG_071204.1KPC-139NG_088399.1 .

[0122] Table 23 below shows the variants of the NDM gene and their NCBI registration numbers.

[0123] Variant typeNCBI No.Variant typeNCBI No.Variant typeNCBI No.NDM-1NG_049326.1NDM-20NG_057455.1NDM-33NG_080782.1NDM-10NG_049327.1NDM-22NG_057612.1NDM-42NG_080783.1NDM-11NG_049328.1NDM-23NG_060570.1NDM-43NG_081701.1NDM-12NG_049329.1NDM-24NG_060571.1NDM-44NG_088409.1NDM-13NG_049330.1NDM-27NG_062358.1NDM-45NG_148636.1NDM-14NG_049331.1NDM-28NG_064729.1NDM-46NG_148637.1NDM-15NG_049332.1NDM-25NG_066711.1NDM-47NG_148638.1NDM-16aNG_049333.1NDM-29NG_067144.1NDM-48NG_148639.1NDM-2NG_049334.1NDM-26NG_067145.1NDM-50NG_149663.1NDM-3NG_049335.1NDM-30NG_071206.1NDM-49NG_149664.1NDM-4NG_049336.1NDM-31NG_071207.1NDM-51NG_157017.1NDM-5NG_049337.1NDM-16bNG_074726.1NDM-52NG_157018.1NDM-6NG_049338.1NDM-36NG_076641.1NDM-53NG_157019.1NDM-7NG_049339.1NDM-37NG_076642.1NDM-54NG_157020.1NDM-8NG_049340.1NDM-34NG_076661.1NDM-55NG_157021.1NDM-9NG_049341.1NDM-35NG_076662.1NDM-56NG_203399.1NDM-17NG_052662.1NDM-38NG_076664.1NDM-57NG_203400.1NDM-18NG_052866.1NDM-39NG_076842.1NDM-58NG_203401.1NDM-19NG_055498.1NDM-40NG_076843.1NDM-60NG_203402.1NDM-21NG_055664.1NDM-41NG_078034.1NDM-61NG_231554.1.

[0124] Table 24 below shows the variants and NCBI registration numbers of OXA-48 family genes.

[0125] Variant typeNCBI No.Variant typeNCBI No.Variant typeNCBI No.OXA-162NG_049461.1OXA-517NG_054666.1OXA-929NG_070750.1OXA-163NG_049462.1OXA-547NG_054693.1OXA-933NG_077983.1OXA-181NG_049482.1OXA-546NG_054959.1OXA-934NG_077984.1OXA-199NG_049495.1OXA-514NG_055475.1OXA-1038NG_078041.1OXA-204NG_049502.1OXA-515NG_055476.1OXA-1039NG_078042.1OXA-232NG_049528.1OXA-519NG_055490.1OXA-1055NG_079243.1OXA-244NG_049539.1OXA-566NG_055499.1OXA-1119NG_081705.1OXA-245NG_049540.1OXA-567NG_055666.1OXA-1146NG_088410.1OXA-247NG_049542.1OXA-535NG_056053.1OXA-1181NG_149665.1OXA-370NG_049661.1OXA-731NG_061613.1OXA-1012NG_203403.1OXA-405NG_049694.1OXA-788NG_064749.1OXA-1200NG_203404.1OXA-416NG_049704.1OXA-793NG_064751.1OXA-1201NG_203405.1OXA-436NG_049722.1OXA-833NG_065443.1OXA-1205NG_203408.1OXA-438NG_049724.1OXA-252NG_066509.1OXA-1207NG_203409.1OXA-439NG_049725.1OXA-894NG_066755.1OXA-1211NG_228675.1OXA-48NG_049762.1OXA-918NG_070182.1OXA-1212NG_228676.1OXA-484NG_049766.1OXA-920NG_070183.1OXA-1213NG_228677.1OXA-505NG_049783.1OXA-923NG_070185.1OXA-1167NG_242303.1OXA-54NG_049794.1OXA-924NG_070186.1 OXA-538NG_052051.1OXA-922NG_070747.1.

[0126] Table 25 below shows the variants of the GES gene and their NCBI registration numbers.

[0127] Variant typeNCBI No.Variant typeNCBI No.Variant typeNCBI No.GES-1NG_049111.1GES-28NG_049131.1GES-41NG_065425.1GES-10NG_049112.1GES-29NG_049132.1GES-42NG_065870.1GES-11NG_049113.1GES-3NG_049133.1GES-43NG_065871.1GES-12NG_049114.1GES-30NG_049134.1GES-44NG_070735.1GES-13NG_049115.1GES-31NG_049135.1GES-45NG_070736.1GES-14NG_049116.1GES-4NG_049136.1GES-46NG_071202.1GES-15NG_049117.1GES-5NG_049137.1GES-47NG_074708.1GES-16NG_049118.1GES-6NG_049138.1GES-48NG_074709.1GES-17NG_049119.1GES-7NG_049139.1GES-49NG_076643.1GES-18NG_049120.1GES-8NG_049140.1GES-50NG_077975.1GES-19NG_049121.1GES-9NG_049141.1GES-51NG_077976.1GES-2NG_049122.1GES-32NG_051512.1GES-52NG_080772.1GES-20NG_049123.1GES-33NG_054709.1GES-53NG_080773.1GES-21NG_049124.1GES-34NG_056177.1GES-54NG_081003.1GES-22NG_049125.1GES-35NG_060520.1GES-55NG_081782.1GES-23NG_049126.1GES-36NG_060521.1GES-56NG_088389.1GES-24NG_049127.1GES-37NG_060522.1GES-57NG_148619.1GES-25NG_049128.1GES-38NG_062215.1GES-58NG_157005.1GES-26NG_049129.1GES-39NG_062216.1GES-59NG_203390.1GES-27NG_049130.1GES-40NG_062356.1.

[0128] 하기 표 26은 VIM-2-like 유전자의 variants 및 NCBI 등록번호이다.

[0129] Variant typeNCBI No.Variant typeNCBI No.Variant typeNCBI No.VIM-53NG_062231.1VIM-41NG_050369.1VIM-66NG_064786.1VIM-10NG_050337.1VIM-44NG_050372.1VIM -65NG_065448.1VIM-11NG_050338.1VIM-45NG_050373.1VIM-67NG_065942.1VIM-15NG_050342.1VIM-46NG_ 050374.1VIM-72NG_070756.1VIM-16NG_050343.1VIM-50NG_050378.1VIM-73NG_070757.1VIM-17NG_050344 .1VIM-51NG_050379.1VIM-74NG_074754.1VIM-18NG_050345.1VIM-6NG_050380.1VIM-75NG_076811.1VIM-2 NG_050347.1VIM-8NG_050382.1VIM-77NG_078035.1VIM-20NG_050348.1VIM-9NG_050383.1VIM-80NG_07926 1.1VIM-23NG_050349.1VIM-56NG_056405.1VIM-82NG_088450.1VIM-24NG_050350.1VIM-58NG_060588.1VIM -81NG_157061.1VIM-3NG_050356.1VIM-60NG_061404.1VIM-84NG_203417.1VIM-30NG_050357.1VIM-62NG_0 63896.1VIM-85NG_203418.1VIM-31NG_050358.1VIM-48NG_064783.1VIM-36NG_050363.1VIM-63NG_064785.1

[0130] Table 27 below shows the variants of the IMP gene and their NCBI registration numbers.

[0131] Variant typeGroupNCBI No.Variant typeGroupNCBI No.IMP-1G1NG_049172.1IMP-21G3NG_049185.1IMP-10G1NG_049173.1IMP-22G3NG_049186.1IMP-25G1NG_049189.1IMP-41G3NG_049205.1IMP-26G1NG_049190.1IMP-44G3NG_049208.1IMP-3G1NG_049194.1IMP-58G3NG_049219.1IMP-30G1NG_049195.1IMP-68G3NG_055584.1IMP-34G1NG_049199.1IMP-93G3NG_079228.1IMP-38G1NG_049202.1IMP-80G4NG_062274.1IMP-4G1NG_049203.1IMP-13G4NG_049176.1IMP-40G1NG_049204.1IMP-17G4NG_049180.1IMP-42G1NG_049206.1IMP-19G4NG_049182.1IMP-52G1NG_049214.1IMP-2G4NG_049183.1IMP-55G1NG_049217.1IMP-20G4NG_049184.1IMP-59G1NG_055477.1IMP-23G4NG_049187.1IMP-6G1NG_049220.1IMP-24G4NG_049188.1IMP-60G1NG_050945.1IMP-27G4NG_049191.1IMP-61G1NG_051166.1IMP-33G4NG_049198.1IMP-66G1NG_054676.1IMP-37G4NG_049201.1IMP-76G1NG_061409.1IMP-46G4NG_064725.1IMP-77G1NG_061410.1IMP-64G4NG_054710.1IMP-78G1NG_061411.1IMP-67G4NG_055271.1IMP-79G1NG_061626.1IMP-8G4NG_049222.1IMP-88G1NG_070737.1IMP-84G4NG_065875.1IMP-89G1NG_070738.1IMP-95G4NG_079887.1IMP-97G1NG_088392.1IMP-96G4NG_080776.1IMP-98G1NG_148620.1IMP-70G5NG_056176.1IMP-69G1NG_055665.1IMP-14G5NG_049177.1IMP-75G1NG_057607.1IMP-18G5NG_049181.1IMP-101G2NG_242291.1IMP-32G5NG_049197.1IMP-102G2NG_231544.1IMP-39G5NG_064724.1IMP-28G2NG_049192.1IMP-48G5NG_049210.1IMP-43G2NG_049207.1IMP-49G5NG_049211.1IMP-5G2NG_049212.1IMP-54G5NG_049216.1IMP-51G2NG_049213.1IMP-56G5NG_049218.1IMP-7G2NG_049221.1IMP-65G5NG_066508.1IMP-81G2NG_065947.1IMP-74G5NG_057606.1IMP-85G2NG_066696.1IMP-83G5NG_065874.1IMP-94G2NG_079229.1IMP-86G5NG_076650.1IMP-15G2NG_049178.1IMP-87G5NG_076651.1IMP-29G2NG_049193.1IMP-91G5NG_076634.1IMP-45G2NG_049209.1IMP-99G5NG_157006.1IMP-53G2NG_049215.1IMP-100G6NG_203391.1IMP-62G2NG_051513.1IMP-12G6NG_049175.1IMP-82G2NG_065873.1IMP-50G6NG_242292.1IMP-9G2NG_049223.1IMP-63G6NG_052049.1IMP-71G2NG_056414.1IMP-90G6NG_074713.1IMP-73G3NG_057463.1IMP-31G7NG_049196.1IMP-11G3NG_049174.1IMP-35G7NG_049200.1IMP-16G3NG_049179.1IMP-92G7NG_079227.1.

[0132] Table 28 below shows representative sequences identified from 10 sequences and 184 variants amplified using KPC primer set1.

[0133] Amplicon SequenceVariant type(SEQ. NO.)GTTCTGCTGTCTTGTCTCTCATGGCCGCTGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTGTGTACGCGATGGATA184KPC 대표서열(KPC_01)(SEQ. 46)GTTATGCTGTCTTGTCTCTCATGGCCGCTGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTGTGTACGCGATGGATA1KPC-147(KPC_02)(SEQ. 47)GTTCTGCTGTCTTGTCTCTCATGGCCGCTGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCACTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTGTGTACGCGATGGATA1KPC-83(KPC-_03)(SEQ. 48)GTTCTGCTGTCTTGTCTCTCATGGCCGCTGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCCGCTCCATCGGTGTGTACGCGATGGATA1KPC-110(KPC_04)(SEQ. 49)GTTCTGCTGTCTTGTCTCTCATGGCCGCTGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTGTGTACGCGATAGATA1KPC-7(KPC_05)(SEQ. 50)GTTCTGCTGTCTTGTCTCTCATGGCCGCTGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACATGACTTTGGCGGCTCCATCGGTGTGTACGCGATGGATA1KPC-119(KPC_06)(SEQ.51)GTTCTGCTGTCTTGTCTCTCATGGCCGCTGTCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTTGTGTACGCGATGGATA1KPC-26(KPC_07)(SEQ. 52)GTTCTGCTGTCTTGTCTYTCATGGCCGCTGGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTGTGTACCGATGGATA1KPC-41(KPC_08) (SEQ. 53)GTTCTGCTGTCTTGTTTCTCATGGCCGCTGGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTTGTGTACGCGATGGATA1KPC-75(KPC_09) (SEQ. 54)ATTCTGCTGTCTTGTCTCTCATGGCCGCTGGCTGGCTTTTCTGCCACCGCGCTGACCAACCTCGTCGCGGAACCATTCGCTAAACTCGAACAGGACTTTGGCGGCTCCATCGGTTGTACGCGATGGATA2KPC-18, 122(KPC_10) (SEQ. 55).

[0134] Table 29 below shows representative sequences identified from 65 sequences and 78 variants amplified using KPC primer set2.

[0135] Amplicon SequenceVariant type(SEQ. NO.)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC78KPC 대표서열(KPC_11)(SEQ. 56)ATACAGTGATAACACCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-179(KPC_12)(SEQ. 57)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCAAACTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-15(KPC_13)(SEQ.58)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-201(KPC_14)(SEQ. 59)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGAACGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-36(KPC_15)(SEQ. 60)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCACTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-187(KPC_16)(SEQ.61)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCCCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-77(KPC_17)(SEQ. 62)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCCGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGCTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-37(KPC_18)(SEQ. 63)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCCGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-118(KPC_19)(SEQ.64)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-150(KPC_20)(SEQ. 65)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGAACTCCGCCATCCCAGGCGATGAGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-197(KPC_21)(SEQ. 66)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-124(KPC_22)(SEQ.67)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-92(KPC_23)(SEQ. 68)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGATGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-12(KPC_24)(SEQ. 69)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCAGAACTCCGCCATCCCAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-208(KPC_25)(SEQ.70)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-115(KPC_26)(SEQ. 71)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCCGAACTCCACCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-48(KPC_27)(SEQ. 72)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGACATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-72(KPC_28)(SEQ.73)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCAATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-168(KPC_29)(SEQ. 74)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCCTACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-191(KPC_30)(SEQ. 75)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGAGACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-159(KPC_31)(SEQ.76)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTATTCCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-206(KPC_32)(SEQ. 77)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-108(KPC_33)(SEQ. 78)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGGC1KPC-131(KPC_34)(SEQ.79)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAGCACGAC1KPC-149(KPC_35)(SEQ. 80)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-107(KPC_36)(SEQ. 81)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGTCGCAGCGGCAGCAGCTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-16(KPC_37)(SEQ.82)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACGAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-43(KPC_38)(SEQ. 83)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-121(KPC_39)(SEQ. 84)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGTTACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-57(KPC_40)(SEQ.85)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGTCGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-143(KPC_41)(SEQ. 86)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCTATACCTCATCGCTGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-194(KPC_42)(SEQ. 87)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCTACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-186(KPC_43)(SEQ.88)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCTATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-88(KPC_44)(SEQ. 89)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCTAGACGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-137(KPC_45)(SEQ. 90)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCTAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-111(KPC_46)(SEQ.91)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-81(KPC_47)(SEQ. 92)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCTATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-13(KPC_48)(SEQ. 93)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTTCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-116(KPC_49)(SEQ.94)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGCACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-129(KPC_50)(SEQ. 95)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGGACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-165(KPC_51)(SEQ. 96)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGGAGCTGAACTCCACCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-127(KPC_52)(SEQ.97)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-196(KPC_53)(SEQ. 98)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGGGCTGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-69(KPC_54)(SEQ. 99)ATACAGTGATAGCGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC1KPC-109(KPC_55)(SEQ.100)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-94, 166(KPC_56)(SEQ. 101)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACCCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-61, 176(KPC_57)(SEQ. 102)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCACCATCCCAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-95, 189(KPC_58)(SEQ.103)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-68, 114(KPC_59)(SEQ. 104)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGCTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-17, 22(KPC_60)(SEQ. 105)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACAGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-49, 99(KPC_61)(SEQ.106)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCACTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-98, 180(KPC_62)(SEQ. 107)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGTCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-85, 126(KPC_63)(SEQ. 108)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCTAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC2KPC-136, 178(KPC_64)(SEQ.109)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCAGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC3KPC-62, 105, 207(KPC_65)(SEQ. 110)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCACCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC3KPC-39, 47, 144(KPC_66)(SEQ. 111)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCAATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC3KPC-51, 140, 170(KPC_67)(SEQ.112)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC3KPC-64, 71, 142(KPC_68)(SEQ. 113)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGCTACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC3KPC-78, 134,125(KPC_69)(SEQ. 114)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCCGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC4KPC-35, 46, 138, 156(KPC_70)(SEQ.115)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGGTACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC4KPC-86, 130, 133, 185(KPC_71)(SEQ. 116)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC5KPC-65, 90, 104, 106, 123(KPC_72)(SEQ. 117)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGGAGCTGGAGCTGAACTCCGCCATCCCAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC5KPC-25, 40, 53, 152, 153(KPC_73)(SEQ.118)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTCTGGACCGCTGGGAGCTGAACTCCGCCATCC CAGGCGATGCGCGCGATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC6KPC-66, 73, 112, 135, 160(KPC_74)(SEQ. 119)ATACAGTGATAACGCCGCCGCCAATTTGTTGCTGAAGGAGTTGGGCGGCCCGGCCGGGCTGACGGCCTTCATGCGCTCTATCGGCGATACCACGTTCCGTTCTGGACCGCTGGGAGCTGGAGCTGAACTCCGCCA TCCCAGGCGATGCGCGCTATACCTCATCGCCGCGCGCCGTGACGGAAAGCTTACAAAAACTGACACTGGGCTCTGCACTGGCTGCGCCGCAGCGGCAGCAGTTTGTTGATTGGCTAAAGGGAAACACGAC15KPC-31, 32, 33, 52, 70, 76, 102, 110, 120, 128, 139, 141, 145, 167, 190 (KPC_75) (SEQ. 120).

[0136] Table 30 below shows representative sequences identified from 19 sequences and 42 variants amplified through NDM primer set1.

[0137] Amplicon SequenceVariant type(SEQ. NO.)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATACTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-50(NDM_01)(SEQ. 121)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCACGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-48(NDM_02)(SEQ. 122)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCAGCCCGACGATTGACCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGAGTTTCGGGGCAGTCACTTCCAACGGTTTGATCGTCAGGGATGG1NDM-10(NDM_03)(SEQ.123)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCACCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-53(NDM_04)(SEQ. 124)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-18(NDM_05)(SEQ. 125)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATATGGTTTTCCGCCAGATCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-34(NDM_06)(SEQ.126)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGAGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-21(NDM_07)(SEQ. 127)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGACAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-33(NDM_08)(SEQ. 128)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGACTGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-56(NDM_09)(SEQ.129)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCTCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-25(NDM_10)(SEQ. 130)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCTCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-54(NDM_11)(SEQ. 131)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCTCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-42(NDM_12)(SEQ.132)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGGCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-2(NDM_13)(SEQ. 133)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCGTGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-38(NDM_14)(SEQ. 134)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGTCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-46(NDM_15)(SEQ.135)TTGCCCAATATTATGCACCCGGTCGCGAAGGTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-41(NDM_16)(SEQ. 136)TTGCGCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-40(NDM_17)(SEQ. 137)TTGCTCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCGACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG1NDM-39(NDM_18)(SEQ.138)TTGCCCAATATTATGCACCCGGTCGCGAAGCTGAGCACCGCATTAGCCGCTGCATTGATGCTGAGCGGTGCATGCCCGGTGAAATCCGCCCGACGATTGGCCAGCAAATGGAAACTGGCG ACCAACGGTTTGGCGATCTGGTTTTCCGCCAGCTCGCACCGAATGTCTGGCAGCACACTTCCTATCTCGACATGCCGGGTTTCGGGGCAGTCGCTTCCAACGGTTTGATCGTCAGGGATGG42Others NDM (NDM_19) (SEQ. 139).

[0138] Table 31 below shows representative sequences identified in four sequences and 51 variants amplified through NDM primer set2.

[0139] Amplicon SequenceVariant typeTGGATCAAGCAGGAGATCAACCTGCCGGTCGCGCTGGCGGTGTGACTCACGCGCATCAGGACAAGATGGGCGGTATGGCCGCGCTGCATGCGGGCGGGGATTGCGACTTATGCCAATGCGTTGTCG2NDM-45, 52(NDM_20) (SEQ. 140)TGGATCAAGCAGGAGATCAACCTGCCGGTCGCGCTGGCGGTGGTGACTCACGCGCATCAGGACAAGATGGGCGGTATGGGCGCGCTGCATGCGGGCGGGGATTGCGACTTATGCCAATGCGTTGTCG3NDM-8, 14, 61(NDM_21) (SEQ. 141)TGGATCAAGCAGGAGATCAACCTGCCGGTCGCGCTGGCGGTGGTGACTCACGCGCATCAGGACAAGATGGGCGGTATGAACGCGCTGCATGCGGGCGGGGATTGCGACTTATGCCAATGCGTTGTCG4NDM-7, 19, 29, 50(NDM_22) (SEQ. 142)TGGATCAAGCAGGAGATCAACCTGCCGGTCGCGCTGGCGGTGGTGACTCACGCGCATCAGGACAAGATGGGCGGTATGGACGCGCTGCATGCGGGCGGGGATTGCGACTTATGCCAATGCGTTGTCG51Other NDM (NDM_23) (SEQ. 143)

[0140] Table 32 below shows representative sequences identified in four sequences and 52 variants amplified through OXA-48 primer set1.

[0141] Amplicon SequenceVariant typeTGGAATGAGAATAAGCAGCAAGGATTTACCAATAATCTTAAACGGGCGAACCAAGCATTTTTACCCGCATCTACATTTAAAATTCCCAATAGCTTGATCGCCCTCGATTTG1OXA-547(OXA_01) (SEQ. 144) TGGAATGAGAATAAGCAGCAAGGATTTACCAATAATCCTAAACGGGCGAACCAAGCATTTTTACCCGCATCTACATTTAAAATTCCCAATAGCTTGATCGCCCTCGATTTG1OXA-918(OXA_02) (SEQ. 145) TGGAATGAGAATAAGCAGCAAGGATTTACCAATAATCCTAAAACGGGCGAACCAAGCATTTTTACCCGCATCTACCTTTAAAATTACCAATAGCTTGATCGCCCTCGATTTG52Other OXA-48 family (OXA_03) (SEQ. 146)

[0142] Table 33 below shows representative sequences identified in 37 sequences and 8 variants amplified by OXA-48 primer set2.

[0143] Amplicon SequenceVariant typeTTCGGCAACGGAGCAAGTCGCATTTCTACGAAAGCTGTATCATAACAAGTTGCATGTATCAGAACGCAGTCAGCGTATCGTCAAGCAAGCCATGCTTACCGAGGCTAATAGTGACTACATAATCCGCGCTAAAACCGGATACTCGACCAGAATTGAGCCTCAGATCGGTTGGTGGGTCGGTTGGGTTGAA1OXA-54(OXA_04)(SEQ. 147)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACTGGATACGAGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-370(OXA_05)(SEQ. 148)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACTGGATACGATACTAAGATTGGTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-567(OXA_06)(SEQ. 149)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACTGGATATGGATACGATACTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-438(OXA_07)(SEQ.150)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACTGGATCCAATACTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-247(OXA_08)(SEQ. 151)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-405(OXA_09)(SEQ. 152)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-517(OXA_10)(SEQ. 153)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGAATCGAACCTAATATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-1212(OXA_11)(SEQ.154)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGAATCGAATCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-920(OXA_12)(SEQ. 155)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-788(OXA_13)(SEQ. 156)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGATCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-1055(OXA_14)(SEQ. 157)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-1200(OXA_15)(SEQ.158)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-1012(OXA_16)(SEQ. 159)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGTATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-1167(OXA_17)(SEQ. 160)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-1146(OXA_18)(SEQ. 161)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGGCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-929(OXA_19)(SEQ.162)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGGCTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-162(OXA_20)(SEQ. 163)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAGGCGAATGCTGATTATATTATTCGGGCGAAAACCGGTTATTCAGGCAGAATTGAGCCGAAATTCGGTTGGTGGGTCGGTTGGGTTGAA1OXA-538(OXA_21)(SEQ. 164)TTCGGCCACTGAGCAAATCACCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-1039(OXA_22)(SEQ. 165)TTCGGCCACTGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATCGTTAAACAAGCCATGCTTACTGAGGCGAATGCTGACTATATTATTCGGGCTAAAACGGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAG1OXA-416(OXA_23)(SEQ.166)TTCGGCCACTGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATAGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA1OXA-515(OXA_24)(SEQ. 167)TTCGGCCATCCAGCAAATCGAATTTTTACGCAAGCTGTACCACAACAAGTTGCATGTCTCGGAGCGTAGTCAGCGTATCGTTAAACAAGCCATGCTGACCGAGGCGAATGCTGATTATATTATTCGGGCGAAAACCGGTTATTCAGTCAGAATTGAGCCGAAAATCGGTTGGTGGGTGGGTTGGGTCGAA1OXA-833(OXA_25)(SEQ. 168)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTCTACCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATTGTCAAACAAGCCATGCTGACCGAGGCAAATGCTGACTATATCATCCGGGCGAAAACTGGCTATTCGGTCAGAATTGAACCGAAAATTGGTTGGTGGGTTGGCTGGGTCGAA1OXA-535(OXA_26)(SEQ. 169)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTAGCATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA1OXA-1205(OXA_27)(SEQ.170)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTGAAATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA1OXA-924(OXA_28)(SEQ. 171)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGATTAGAATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA1OXA-1119(OXA_29)(SEQ. 172)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTTGACTAGAATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA1OXA-1211(OXA_30)(SEQ. 173)TTCGGCTATCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTGGAATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA1OXA-922(OXA_31)(SEQ.174)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTGGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA2OXA-244, 1213(OXA_32)(SEQ. 175)TTCGGCCACTGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA2OXA-204, 514(OXA_33)(SEQ. 176)TTCGGCCACTGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA2OXA-199, 894(OXA_34)(SEQ. 177)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTGGAATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA2OXA-484, 1207(OXA_35)(SEQ.178)TTCGGCCACGGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACTGGATACGATACTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA3OXA-163, 439, 731(OXA_36)(SEQ. 179)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA3OXA-181, 923, 1181(OXA_37)(SEQ. 180)TTCGGCTACCCAGCAAATCGCTTTTTTACGCAAGCTGTATCACAACAAGCTGCACGTTTCTGAGCGTAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACGGGATACTCGACTAGTATCGAACCTAAGATTGGCTGGTGGGTTGGTTGGGTTGAA3OXA-232, 793, 1201(OXA_38)(SEQ. 181)TTCGGCCATTGAGCAAATCAGCTTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGTCAGCGCATCGTGAAACAAGCCATGCTGACCGAAGCCAATGGCGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA4OXA-252, 546, 547, 1038(OXA_39)(SEQ.182)TTCGGCCACGGAGCAAATCAGCTTTTAAGAAAGCTGTATCACAATAAGTTACACGTATCGGAGCGCAGCCAGCGTATTGTCAAACAAGCCATGCTGACCGAAGCCAATGGTGACTATATTATTCGGGCTAAAACTGGATACTCGACTAGAATCGAACCTAAGATTGGCTGGTGGGTCGGTTGGGTTGAA8OXA-48, 245, 505, 519, 566, 918, 933, 934(OXA_40) (SEQ. 183).

[0144] Table 34 below shows representative sequences identified from 25 sequences and 18 variants amplified by GES primer set1 and mix1.

[0145] Amplicon Sequence (mix1의 경우 추가)Variant typeCTGCAATGACGCAGTATTTTCGCAAAATTGGCGACTCTGTGAGTAGGCTAGACCGGAAAGAGCCGGAGATGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACTTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACATTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAAAAA(A)1GES-44(GES_01)(SEQ. 184)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGACGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-37(GES_02)(SEQ. 185)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCAGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-27(GES_03)(SEQ.186)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGAAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-36(GES_04)(SEQ. 187)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAACGACAACACACCTGGCGACCTCAGAGATACAACCACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGCCCACCTCGACCCACACAATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACATTACGAGCGGGTTTTCCTAAAGATTGGGTTATTGGAGAGAAA(A)1GES-59(GES_05)(SEQ. 188)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAAATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-28(GES_06)(SEQ.189)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACCACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGCCCACCTCGACCCACACAATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACATTACGAGCGGGTTTTCCTAAAGATTGGGTTATTGGAGAGAAA(A)1GES-41(GES_07)(SEQ. 190)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCAAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-25(GES_08)(SEQ. 191)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAAAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-21(GES_09)(SEQ.192)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACGCTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-16(GES_10)(SEQ. 193)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGGGCGACAACACACCTGGCGACCTCAGAGATACAACCACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGCCCACCTCGACCCACACAATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACATTACGAGCGGGTTTTCCTAAAGATTGGGTTATTGGAGAGAAA(A)1GES-32(GES_11)(SEQ. 194)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCCAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-9(GES_12)(SEQ.195)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACGCTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-31(GES_13)(SEQ. 196)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACTTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-1(GES_14)(SEQ. 197)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGCTGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-22(GES_15)(SEQ.198)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGTTGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAATCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-35(GES_16)(SEQ. 199)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-42(GES_17)(SEQ. 200)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGGCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-29(GES_18)(SEQ.201)CTGCAATGACGCAGTATTTTCGTAAAATTGGGGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-18(GES_19)(SEQ. 202)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAATGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)1GES-34(GES_20)(SEQ. 203)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAACGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)2GES-2, 13(GES_21)(SEQ.204)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAATGAGCCGGAGATGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)2GES-46, 57(GES_22)(SEQ. 205)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGTCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)3GES-15, 43, 55(GES_23)(SEQ. 206)CTGCAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGGGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)13GES-3, 7, 8, 10, 11, 12, 17, 19, 26, 38, 45, 52, 56(GES_24)(SEQ.207)CTGCAAATGACGCAGTATTTTCGTAAAATTGGCGACTCTGTGAGTCGGCTAGACCGGAAAGAGCCGGAGATGAGCGACAACACACCTGGCGACCTCAGAGATACAACTACGCCTATTGCTATGGCACGTACTGTGGC TAAAGTCCTCTATGGCGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAA(A)18GES-4; 5, 6, 14, 20, 24, 30, 33, 39, 40, 47, 48, 49, 50, 51, 53, 54, 58 (GES_25) (SEQ. 208).

[0146] Table 35 below shows representative sequences identified in 9 sequences and 47 variants amplified by GES primer set2 and mix2.

[0147] Amplicon Sequence (set2의 경우 추가)Variant typeCGTACTGTGGCAAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAAA(A)1GES-25(GES_26)(SEQ. 209)CGTACTGTGGCCAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAAA(A)1GES-9(GES_27)(SEQ. 210)CGTACTGTGGCTAAAATCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAAA(A)1GES-35(GES_28)(SEQ. 211)CGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAAAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAAA(A)1GES-21(GES_29)(SEQ. 212)CGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACTTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAGAAAA(A)1GES-1(GES_30)(SEQ.213) CGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACTTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACATTACGAGCGGGTTTTCCTAAAGATTGGGTTGTTGGAGAAAAAA(A)1GES-44(GES_31) (SEQ. 214) CGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACGCTACGAGCGGGTTTTCCTAAAGATTGGGTTGGAGAGAAAA(A)2GES-16, 31(GES_32) (SEQ. 215) CGTACTGTGGCTAAAGTCCTCTATGGCGGGCGCACTGACGCCCACCTCGACCCACACAATTGAGAGGTGGCTGATCGGAAACCAAACGGGAGACGCGACATTACGAGCGGGTTTTCCTAAAGATTGGGTTATTGGAGAGAAAA(A)3GES-32, 41, 59(GES_33) (SEQ. 216) CGTACTGTGGCTAAAGTCCTCTATGGCGGCGCACTGACGTCCACCTCGACCCACACCATTGAGAGGTGGCTGATCGGAAACCAAACGGGGAGACGCGACACTACGAGCGGGTTTTCCTAAAGATTGGGTTGGAGAGAAAA(A)47Other GES (GES_34) (SEQ. 217).

[0148] Table 36 below shows representative sequences identified in 7 sequences and 30 variants amplified through VIM primer set1.

[0149] Amplicon SequenceVariant typeCGACAGTCAACGAAATTCCGGTCGGGGAGGTCCGGCTTTACCAGATTGCCGATGGTGTTTGGTCGCATATCGCAACGCAGTCGTTTGATGGCGCAGTCTACCCGTCCAATGGTCTCATTGTCCGTGATGGTGATGAGTTGCTTTTGATTG1VIM-30(VIM_01)(SEQ. 218)CGACAGTCAGCGAAATTCCGGTCGGGGAGGTCCGGCTTTACCAGATTGCCGATGGTGTTTGGTCGCATATCGCAACGAAGTCGTTTGATGGCGCAGTCTACCCGTCCAATGGTCTCATTGTCCGTGATGGTGATGAGTTGCTTTTGATTG1VIM-3(VIM_02)(SEQ. 219)CGACAGTCAGCGAAATTCCGGTCGGGGAGGTCCGGCTTTACCAGATTGCCGATGGTGTTTGGTTGCATATCGCAACGCAGTCGTTTGATGGCGCAGTCTACCCGTCCAATGGTCTCATTGTCCGTGATGGTGATGAGTTGCTTTTGATTG1VIM-16(VIM_03)(SEQ. 220)CGATAGTCAGCGAAATTCCGGTCGGGGAGGTCCGGCTTTACCAGATTGCCGATGGTGTTTGGTCGCATATCGCAACGCAGTCGTTTGATGGCGCAGTCTACCCGTCCAATGGTCTCATTGTCCGTGATGGTGATGAGTTGCTTTTGATTG1VIM-45(VIM_04)(SEQ. 221)CGACAGCCAGCGAAATTCCGGTCGGGGAGGTCCGGCTTTACCAGATTGCCGATGGTGTTTGGTCGCATATCGCAACGCAGTCGTTTGATGGCGCAGTCTACCCGTCCAATGGTCTCATTGTCCGTGATGGTGATGAGTTGCTTTTGATTG3VIM-46, 65, 67(VIM_05)(SEQ.222) CGACAGTCAGCGAAATTCCGGTCGGGGAGGTCCGGCTTTACCAGATTGCCGATGGTGTTTGGTCGCATATCGCAACGCGGTCGTTTGATGGCGCAGTCTACCCGTCCAATGGTCTCATTGTCCGTGATGGTGATGAGTTGCTTTTGATTG6VIM-6, 18, 36, 82, 84, 85 (VIM_06) (SEQ. 223) CGACAGTCAGCGAAATTCCGGTCGGGGAGGTCCGGCTTTACCAGATTGCCGATGGTGTTTGGTCGCATATCGCAACGCAGTCGTTTGATGGCGCAGTCTACCCGTCCAATGGTCTCATTGTCCGTGATGGTGATGAGTTGCTTTTGATTG30Other VIM-2-like (VIM_07) (SEQ. 224).

[0150] Table 37 below shows representative sequences identified in 15 sequences and 26 variants amplified using VIM primer set2.

[0151] Amplicon SequenceVariant typeTGCAGTCTCCACGCACTTTCATAACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-85(VIM_08)(SEQ. 225)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGCTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-75(VIM_09)(SEQ. 226)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-80(VIM_10)(SEQ.227)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTCATGAGT1VIM-31(VIM_11)(SEQ. 228)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTTTGGTGGTTGTGCGATTTATGAGT1VIM-15(VIM_12)(SEQ. 229)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGGGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-74(VIM_13)(SEQ.230)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTTCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-73(VIM_14)(SEQ. 231)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGGACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-77(VIM_15)(SEQ. 232)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-18(VIM_16)(SEQ.233)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGATACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-9(VIM_17)(SEQ. 234)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGGCACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCGGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-8(VIM_18)(SEQ. 235)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTTGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-66(VIM_19)(SEQ.236)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCGCCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT1VIM-44(VIM_20)(SEQ. 237)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAGCGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT4VIM-3, 6, 11, 50(VIM_21)(SEQ. 238)TGCAGTCTCCACGCACTTTCATGACGACCGCGTCGGCGGCGTTGATGTCCTTCGGGCGGCTGGGGTGGCAACGTACGCATCACCGTCGACACGCCGGCTAGCCGAGGTAGAGGGGAACGAGATTCCCACGCACTCTCTAGAAGGACTCTCATCGAGCGGGGACGCAGTGCGCTTCGGTCCAGTAGAACTCTTCTATCCTGGTGCTGCGCATTCGACCGACAACTTAGTTGTGTACGTCCCGTCTGCGAGTGTGCTCTATGGTGGTTGTGCGATTTATGAGT26그외 VIM-2-like(VIM_22)(SEQ. 239).

[0152] Table 38 below shows representative sequences identified in 11 sequences and 12 variants amplified through IMP primer set1.

[0153] Amplicon SequenceVariant typeGCTTAATTCTCAATCCATCCCCACGTATGCGTCTGAATTAACTAATGAGCTGCTTAAAAAAGACGGTAAGGTTCAAGCTAAAAATTCATTTGGCGGGGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCAGGACACACTCCAGATAACCTAGTAGTTTGGCTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTCTAGGTTATTTGGGTGACGCAAATTTAGAAGC1IMP-59(IMP_01)(SEQ. 240)GCTTAATTCTCAATCTATTCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTGCAAGCTAAACACTCATTTAGCGGAGTTAGTTATTGGCTAGTCAAAAATAAAATTGAAGTTTTTTATCCTGGCCCAGGCCACACTCAAGATAACGTAGTGGTTTGGTTACCTGAAAATAAAATTTTATTCGGTGGTTGCTTTGTTAAACCTCACGGTCTTGGTAATTTGGGTGACGCAAATTTAGAAGC1IMP-79(IMP_02)(SEQ. 241)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGCAAGGTTCAAGCTACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGGGTGACGCAAATATAGAAGC1IMP-69(IMP_03)(SEQ.242)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCAAAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGGGTGACGCAAATATAGAAGC1IMP-89(IMP_04)(SEQ. 243)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTAAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGGGTGACGCAAATATAGAAGC1IMP-60(IMP_05)(SEQ. 244)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGAGTGACGCAAATATAGAAGC1IMP-25(IMP_06)(SEQ.245)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGAAAATATTTTTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAAATTGGGTGACGCAAATATAGAAGC1IMP-55(IMP_07)(SEQ. 246)GCTTATTTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGGGTGACGCAAATATAGAAGC1IMP-61(IMP_08)(SEQ. 247)GCTTAATTCTCAATCCATCCCCACGTATGCGTCTGAATTAACTAATGAGCTGCTTAAAAAAGACGGTAAGGTTCAAGCTAAAAATTCATTTGGCGGGGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCAGGACACACTCCAGATAACCTAGTAGTTTGGCTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTCTAGGTAATTTGGGTGACGCAAATTTAGAAGC4IMP-4, 26, 38, 88(IMP_09)(SEQ.248)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTAAAAATAAAATTGAAGT TTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGGGTGACGCAAATATAGAAGC4IMP-3, 34, 78, 97(IMP_10)(SEQ. 249)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTAAAAATAAAATTGAAGT TTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGGGTGACGCAAATATAGAAGC12IMP-1, 6, 10, 30, 40, 42, 52, 66, 75, 76, 77, 98 (IMP_11) (SEQ. 250).

[0154] A total of 43 variants can be detected in the G2~G7 group using IMP primer set 1, including 17 G2, 14 G4, 8 G5, 1 G6, and 3 G7, excluding IMP-29. The sequences identified according to variant are shown in Table 39 below.

[0155] Amplicon Sequence (G2~G7 group)Variant typeGCTTAATTCTCAATCTATCCCCACATATGCATCTAAATTAACAAATGAGCTTCTTAAAAAGAACGGTAATGCGCAAGCCGTAAACTCATTTAGTGGCGTTAGCTATTGGCTAGTTAAACATAAATTGAAGTTTTCTA TCCAGGACCAGGGCACACTCAGGATAATGTAGTGGTTTGGTTGCCTGAAAAGAAAATTTTTTGGCGGTTGTTTTATTAAGCCGGACGGTCTTGGTTATTTGGGAGACGCAAATCTAGAAGC1IMP-92(IMP_12)(SEQ. 251)GCTTAATTCTCAATCTATCCCCACATATGCTTCTGAATTAACAAATGAACTTCTTAAAAAGACAATAAGGTACAAGCTAAACTCTTTTTATGGGGTTAGTTATTCACTAATAAAAAACAAATTTGAAGTTTTTTA TCCAGGCCCAGGGCACACTCAAGATAACGTAGTGGTTTGGTTACCTGAAAAGAAAATTTTATTCGGTGGTTGCTTTGTTAAACCGGACGGTCTTGGCTATTTGGGGGACGCAAATTTAGAAGC1IMP-32(IMP_13)(SEQ. 252)GCTTAATTCTCAATCTATCCCCACGTATGCATCAGAATTAACAAATGAACTCCTTAAAAAAGACGGTAAGGTACAAGCTAAATTCATTTAGCGGAGTTAGTTATTGGCTAGTTAAGAATAAGGTTGAAATTTTTTA TCCTGGTCCTGGGCACACTCCAGATAACGTAGTGGTTTGGTTACCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTAAACCGTACGGTCTTGGTAATTTGGGTGATGCAAATTTAGAAGC1IMP-85(IMP_14)(SEQ.253)GCTTAATTCTCAATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAAGTACAAGCTAAAAATTCATTTAGCGGAGCTAGCTATTGGCTAGTTAAGAAAAAGATTGAAGTTTTTTATCCTGGTCCAGGGCACACTCCAGATAACGTAGTGGTTTGGCTACCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTTAAACCGTACGGTCTAGGTAATTTGGGTGACGCAAATTTAGAAGC1IMP-101(IMP_15)(SEQ. 254)GCTTAATTCTCAATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTACAAGCTAAAAATTCATTTAGCGGAGTTAGCTATTGGCTAGTTAAGAAAAAGATTGAAGTTTTTTA TCCTGTCCAGGGCACGCTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTTAAACCCTACGGTCTAGGTAATTTGGGTGACGCCAATTTAGAAGC1IMP-82(IMP_16)(SEQ. 255)GCTTAATTCTCAATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTACAAGCTAAATATTCATTTAGCGGAGTTAGCTATTGGCTAGTTAAGAAAAAGATTGAAGTTTTTTATCCTGGTTCAGGGCACGCTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTTAAACCCTACGGTCTAGGTAATTTGGGTGACGCAAATTTAGAAGC1IMP-53(IMP_17)(SEQ.256)GCTTAATTCTCAATCTATTCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAGAACGGTAAGGTGCAAGCTAAAAACTCATTTAGCGGAGTTAGTTATTGGCTAGTTAAAAATAAAATTGAAATTTTTTAT CCCGGTCCGGGGCACACTCAAGATAACGTAGTGGTTTGGCTACCTGAAAACAAAATTTTATTCGGTGGTTGTTTTGTTAAACCGGACGGTCTTGGTAATTTGGATGACGCAAATTTAGAAGC1IMP-100(IMP_18)(SEQ. 257)GCTTAATTCTCAATCTATTCCCACGTATGCATCTGAATTAACAAGAAATGAACTTTTGAAAAAATCCGGTAAGGTACAAGCTAAATATTCATTTAGCGAAGTTAGCTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCTGGCCCAGGTCACACTCAAGATAACCTAGTGGTTTGGTTGCCTGAAAGTAAAATTTTATTCGGTGGTTGCTTTGTTAAACCTCACGGTCTTGGCAATTTAGGTGACGCAAATTTAGAAGC1IMP-33(IMP_19)(SEQ. 258)GCTTAATTCTCAATCTATTCCCACGTATGCATCTGAATTAACAAGAAATGAACTTTTGAAAAAATCCGGTAAGGTACAAGCTAAATATTCATTTAGCGGAGTTAGCTATTGGCTAGTTAAAAATAAAATTGAAGTTTTCTACCCTGGCCCAGGTCACACTCAAGATAACCTAGTGGTTTGGTTGCCTGAAAGTGAAATTTTATTCGGTGGTTGCTTTATTAAACCTCACGGTCTTGGCAATTTAGGTGACGCAAATTTAGAAGC1IMP-17(IMP_20)(SEQ.259)GCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAACACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGAAAATATTATTCGGTGGTTGTTTTATTAAACCGTACGGTTTAGGCAATTTGGGTGACGCAAATATAGAAGC1IMP-80(IMP_21)(SEQ. 260)GCTTAATTCTCAATCTATCCCCACATATGCATCTAAATTAACAAATGAGCTTCTTAAAAAGAACGGTAATGCGCAAGCCGAAAACTCATTTAGTGGCGTTAGCTATTGGCTAGTTAAACATAAAATTGAAGTTTTCTATCCAGGACCAGGGCACACTCAGGATAATGTAGTGGTTTGGTTGCCTGAAAAGAAAATTTTATTTGGCGGTTGTTTTATTAAGCCGGACGGTCTTGGTTATTTGGGAGACGCAAATCTAGAAGC2IMP-31, 35(IMP_22)(SEQ. 261)GCTTAATTCTCAATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAAGTACAAGCTAAAAATTCATTTAGCGGAGCTAGCTATTGGCTAGTTAAGAAAAAGATTGAAGTTTTTTATCCTGGTCCAGGGCACACTCCAGATAACGTAGTGGTTTGGCTACCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTTAAACCGTACGGTCTAGGTAATTTGGGTGACGCAAATGTAGAAGC2IMP-5, 81(IMP_23)(SEQ.262)GCTTAATTCTCAATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTACAAGCTAAAAATTCATTTAGCGGAGGTAGCTATTGGCTAGTTAATAATAAGATTGAAGTTTTTTATCCTGGTCCAGGGCACACTCCAGATAACGTAGTGGTTTGGCTACCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTTAAACCGTACGGTCTTGGTAATTTGGGTGACGCAAATTTAGAAGC2IMP-15, 62(IMP_24)(SEQ. 263)GCTTAATTCTCAATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTACAAGCTAAATATTCATTTAGCGGAGTTAGCTATTGGCTAGTTAAGAAAAAGATTGAAGT TTTTTATCCTGGTCCAGGGCACGCTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTTAAACCCTACGGTCTAGGTAATTTGGGTGACGCAAATTTAGAAGC2IMP-9, 45(IMP_25)(SEQ. 264)GCTTAATTCTCAATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTACAAGCTAAAAATTCATTTGGCGGAGTTAGCTATTGGCTAGTTAAGAATAAGATTGAAGTTTTTTATCCTGGTCCAGGGCACACTCCAGATAACGTAGTGGTTTGGCTACCTGAAAATAGAGTTTTGTTCGGTGGTTGTTTTGTTAAACCGTACGGTCTTGGTAATTTGGGTGACGCAAATTTAGAAGC3IMP-28, 94, 102(IMP_26)(SEQ.265)GCTTAATTCTCAATCTATTCCCACGTATGCATCTGAATTAACAAATGAACTTTTGAAAAAATCCGGTAAGGTACAAGCTAAATATTCATTTAGCGAAGTTAGCTATTGGCTAGTTAAAAATAAAATTGAAGTTTTCTACCCTGGCCCAGGTCACACTCAAGATAACCTAGTGGTTTGGTTGCCTGAAAGTAAAATTTTATTCGGTGGTTGCTTTATTAAACCTCACGGTCTTGGCAATTTAGGTGACGCAAATTTAGAAGC3IMP-13, 37, 84(IMP_27)(SEQ. 266)GCTTAATTCTCAATCTATTCCAACATATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTACAAGCTAAAAATTCATTTAGCGGAGCCAGCTATTGGTTAGTTAAGAAAAAGATTGAAAT TTTTTATCCTGGCCCAGGGCACACTCCAGATAACGTAGTGGTTTGGCTACCTGAACATAGAGTTTTGTTTGGTGGTTGTTTTGTTAAACCGTATGGTCTAGGTAATTTGGGTGACGCAAATTTAGAAGC4IMP-7, 43, 51, 71(IMP_28)(SEQ. 267)GCTTAATTCTCAATCTATCCCCACATATGCTTCTGAATTAACAATGAACTTCTTAAAAAAGACAATAAGGTACAAGCTAAACACTCTTTTAATGGGGTTAGTTATTCACTAATTAAAAACAAAATTGAAGTTTTTTATCCAGGCCCAGGGCACACTCAAGATAACGTAGTGGTTTGGTTACCTGAAAAGAAAATTTTATTCGGTGGTTGCTTTGTTAAACCGGACGGTCTTGGCTATTTGGGGGACGCAAATTTAGAAGC7IMP-14, 48, 54, 65, 86, 87, 91(IMP_29)(SEQ.268)GCTTAATTCTCAATCTATTCCCACGTATGCATCTGAATTAACAAATGAACTTCTTAAAAAAGACGGTAAGGTGCAAGCTAAAAACTCATTTAGCGGAGTTAGTTATTGGCTAGTTTAAAAATAAAATTGAAGT TTTTTATCCCGGCCCGGGGCACACTCAAGATAACGTAGTGGTTTGGTTACCTGAAAAGAAAATTTTATTCGGTGGTTGTTTTGTTAAACCGGACGGTCTTGGTAATTTGGGTGACGCAAATTTAGAAGC8IMP-2, 8, 19, 20, 23, 24, 46, 96 (IMP_30) (SEQ. 269).

[0156] Table 40 below shows representative sequences identified in 9 and 14 variants of the IMP G1 group amplified using IMP primer set 2. Variants of the IMP G2-G7 groups were not detected.

[0157] Amplicon SequenceVariant typeTCATTTTCATAGCGACAGCACGGGCGGAATAGAGTGGCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTAAAAGGA1IMP-60(IMP_31)(SEQ. 270)TCATTTTCATAGCGACAGCACGGGCGGAATAGAGTGGCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAACACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGA1IMP-88(IMP_32)(SEQ. 271)TCATTTTCATAGCGACAGCACGGGCGGAATAGAGTGGCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGCAAGGTTCAAGCTACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAGGA1IMP-69(IMP_33)(SEQ.272)TCATTTTCATAGCGACAGCACGGGCGGAATAGAGTGGCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCAAAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGA1IMP-97(IMP_34)(SEQ. 273)TCATTTTCATAGCGACAGCACGGGCGGAATAGAGTGGCTTATTTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGA1IMP-61(IMP_35)(SEQ. 274)TCATTTTCATAGCGACAGCACGGGCGGAATAGAGTGGCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAGGA2IMP-78, 98(IMP_36)(SEQ.275)TCATTTTCATAGCGACAGCACGGGCGGAATAGGGTGGCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGCTGCCTGAAAGGA2IMP-3, 34(IMP_37)(SEQ. 276)TCATTTTCATAGTGACAGCACGGGCGGAATAGAGTGGCTTAATTCTCAATCCATCCCCACGTATGCGTCTGAATTAACTAATGAGCTGCTTAAAAAAGACGGTAAGGTTCAAGCTAAAAATTCATTTGGCGGGGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCAGGACACACTCCAGATAACCTAGTAGTTTGGCTGCCTGAAAGGA5IMP-4, 26, 38, 59, 89(IMP_38)(SEQ. 277)TCATTTTCATAGCGACAGCACGGGCGGAATAGAGTGGCTTAATTCTCGATCTATCCCCACGTATGCATCTGAATTAACAAATGAACTGCTTAAAAAAGACGGTAAGGTTCAAGCCACAAATTCATTTAGCGGAGTTAACTATTGGCTAGTTAAAAATAAAATTGAAGTTTTTTATCCAGGCCCGGGACACACTCCAGATAACGTAGTGGTTTGGTTGCCTGAAAGGA14IMP-1, 10, 25, 30, 40, 42, 52, 55, 66, 75, 76, 77, 79(IMP_39)(SEQ. 278).

[0158] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions merely represent preferred embodiments and are not intended to limit the scope of the present invention. In other words, the substantial scope of the present invention is defined by the appended claims and their equivalents.

[0159] Numerical ranges are inclusive of the values ​​defined in the ranges above. Any maximum numerical limitation given throughout this specification includes any lower numerical limitation, as if that lower numerical limitation were explicitly stated. Any minimum numerical limitation given throughout this specification includes any higher numerical limitation, as if that higher numerical limitation were explicitly stated. Any numerical limitation given throughout this specification will include any better numerical range within the broader numerical range, as if that narrower numerical limitation were explicitly stated.

Claims

1. A first primer set consisting of a forward primer represented by sequence number 1 or sequence number 3 and a reverse primer represented by sequence number 2 or sequence number 4; A second primer set comprising a forward primer represented by SEQ ID NO: 5 or SEQ ID NO: 7 and a reverse primer represented by SEQ ID NO: 6 or SEQ ID NO: 8; A third primer set consisting of a forward primer represented by SEQ ID NO: 9 or SEQ ID NO: 11 and a reverse primer represented by SEQ ID NO: 10 or SEQ ID NO: 12; A fourth primer set consisting of a forward primer represented by SEQ ID NO: 13 or SEQ ID NO: 15 and a reverse primer represented by SEQ ID NO: 14 or SEQ ID NO: 16; A fifth primer set consisting of a forward primer represented by SEQ ID NO: 17 or SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 18 or SEQ ID NO: 20; and A composition for detecting carbapenemase producing Enterobacterales (CPE), comprising at least one primer set selected from the group consisting of a forward primer represented by SEQ ID NO: 21 or SEQ ID NO: 23 and a sixth primer set consisting of a reverse primer represented by SEQ ID NO: 22 or SEQ ID NO:

24.

2. In paragraph 1, The first primer set specifically detects the KPC gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 46 to SEQ ID NO: 120, The second primer set specifically detects the NDM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 121 to SEQ ID NO: 143, The third primer set specifically detects the OXA-48 gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 144 to SEQ ID NO: 183, The fourth primer set specifically detects a GES gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 184 to SEQ ID NO: 217, The fifth primer set specifically detects the VIM gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 218 to SEQ ID NO: 239, A composition characterized in that the sixth primer set specifically detects an IMP gene or a variant thereof comprising any one of the base sequences of SEQ ID NO: 240 to SEQ ID NO:

278.

3. A composition according to claim 1, characterized in that the composition is for a single polymerase chain reaction (single PCR), a triplex PCR, a multiplex PCR, a real-time polymerase chain reaction (real time PCR), a triplex real-time PCR, or a multiplex real-time PCR.

4. In paragraph 1, The above first primer set is composed of a forward primer represented by sequence number 1 and a reverse primer represented by sequence number 2, The second primer set is composed of a forward primer represented by sequence number 5 and a reverse primer represented by sequence number 6, The third primer set above is composed of a forward primer represented by SEQ ID NO: 9 and a reverse primer represented by SEQ ID NO: 10, The fourth primer set above is composed of a forward primer represented by SEQ ID NO: 13 and a reverse primer represented by SEQ ID NO: 16, The fifth primer set above is composed of a forward primer represented by SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 20, A composition characterized in that the sixth primer set is composed of a forward primer represented by SEQ ID NO: 21 and a reverse primer represented by SEQ ID NO:

22.

5. In the fourth paragraph, the composition is characterized in that it is for triplex PCR or triplex real time PCR, comprising the first primer set, the second primer set, and the third primer set.

6. A composition according to claim 5, characterized in that the composition further comprises a first probe represented by SEQ ID NO: 27, a second probe represented by SEQ ID NO: 28, and a third probe represented by SEQ ID NO:

29.

7. In the fourth paragraph, the composition is characterized in that it is for triplex PCR or triplex real time PCR, comprising the fourth primer set, the fifth primer set, and the sixth primer set.

8. A composition according to claim 7, characterized in that the composition further comprises a fourth probe represented by SEQ ID NO: 30, a fifth probe represented by SEQ ID NO: 31, and a sixth probe represented by SEQ ID NO:

32.

9. Step of isolating DNA from the sample (Step 1); The DNA separated in the above first step is used as a template A first primer set consisting of a forward primer represented by sequence number 1 and a reverse primer represented by sequence number 2, A second primer set consisting of a forward primer represented by sequence number 5 and a reverse primer represented by sequence number 6, A third primer set consisting of a forward primer represented by sequence number 9 and a reverse primer represented by sequence number 10; A fourth primer set consisting of a forward primer represented by sequence number 13 and a reverse primer represented by sequence number 16; A fifth primer set consisting of a forward primer represented by SEQ ID NO: 19 and a reverse primer represented by SEQ ID NO: 20, and A step (second step) of performing a polymerase chain reaction (PCR) using at least one primer set selected from the group consisting of a forward primer represented by sequence number 21 and a reverse primer represented by sequence number 22; A step (step 3) of performing electrophoresis on the PCR amplification product obtained in the above step 2; and A method for detecting a carbapenemase-producing Enterobacterales (CPE) strain, comprising a step (step 4) of detecting a carbapenemase-producing Enterobacterales (CPE) strain based on the presence or absence of a PCR amplification product confirmed in the above-mentioned step 3.

10. In the 9th paragraph, in the 4th step, If a band with a size of 130 bp exists in the above PCR amplification product, it is determined that a CPE strain having the KPC gene or a variant thereof exists in the sample. If a band with a size of 111 bp exists in the PCR amplification product, it is determined that a CPE strain having the OXA-48 gene or a variant thereof exists in the sample. If a band with a size of 242 bp exists in the PCR amplification product, it is determined that a CPE strain having the NDM gene or a variant thereof exists in the sample. If a band with a size of 267 bp exists in the PCR amplification product, it is determined that a CPE strain having the GES gene or a variant thereof exists in the sample. If a band with a size of 281 bp is present in the PCR amplification product, it is determined that a CPE strain having a VIM-2-like gene or a variant thereof is present in the sample. A method characterized in that if a band having a size of 261 bp is present in the PCR amplification product, it is determined that a CPE strain having the IMP G1 gene or a variant thereof is present in the sample.

11. In the 9th paragraph, in the second step, A method characterized in that a triplex PCR or triplex real time PCR is performed using the first primer set, the second primer set, and the third primer set.

12. In the 11th paragraph, when performing triplex real time PCR in the second step, A method characterized by additionally using a first probe represented by sequence number 27, a second probe represented by sequence number 28, and a third probe represented by sequence number 29.

13. In the 9th paragraph, in the second step, A method characterized in that a triplex PCR or triplex real time PCR is performed using the fourth primer set, the fifth primer set, and the sixth primer set.

14. In the 13th paragraph, when performing triplex real time PCR in the second step, A method characterized by additionally using a fourth probe represented by sequence number 30, a fifth probe represented by sequence number 31, and a sixth probe represented by sequence number 32.

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