메뉴 건너뛰기




Volumn 6, Issue 7, 2012, Pages

A simple strain typing assay for Trypanosoma cruzi: Discrimination of major evolutionary lineages from a single amplification product

Author keywords

[No Author keywords available]

Indexed keywords

C 5 STEROL DESATURASE; OXYGENASE; UNCLASSIFIED DRUG; PROTOZOAL DNA; RESTRICTION ENDONUCLEASE;

EID: 84864622808     PISSN: 19352727     EISSN: 19352735     Source Type: Journal    
DOI: 10.1371/journal.pntd.0001777     Document Type: Article
Times cited : (71)

References (45)
  • 2
    • 0021118973 scopus 로고
    • The natural heterogeneity of Trypanosoma cruzi: biological and medical implications
    • Dvorak JA, ((1984)) The natural heterogeneity of Trypanosoma cruzi: biological and medical implications. J Cell Biochem 24:: 357--371.
    • (1984) J Cell Biochem , vol.24 , pp. 357-371
    • Dvorak, J.A.1
  • 3
    • 0000595112 scopus 로고    scopus 로고
    • Isozyme variability in Trypanosoma cruzi, the agent of Chagas' disease: Genetical, taxomonical, and epidemiological significance
    • Tibayrenc M, Ayala FJ, ((1998)) Isozyme variability in Trypanosoma cruzi, the agent of Chagas' disease: Genetical, taxomonical, and epidemiological significance. Evolution 42:: 277--292.
    • (1998) Evolution , vol.42 , pp. 277-292
    • Tibayrenc, M.1    Ayala, F.J.2
  • 4
    • 0342636508 scopus 로고
    • Natural populations of Trypanosoma cruzi, the agent of Chagas disease, have a complex multiclonal structure
    • Tibayrenc M, Ward P, Moya A, Ayala FJ, ((1986)) Natural populations of Trypanosoma cruzi, the agent of Chagas disease, have a complex multiclonal structure. Proc Natl Acad Sci U S A 83:: 115--119.
    • (1986) Proc Natl Acad Sci U S A , vol.83 , pp. 115-119
    • Tibayrenc, M.1    Ward, P.2    Moya, A.3    Ayala, F.J.4
  • 5
    • 0036746010 scopus 로고    scopus 로고
    • The clonal theory of parasitic protozoa: 12 years on
    • Tibayrenc M, Ayala FJ, ((2002)) The clonal theory of parasitic protozoa: 12 years on. Trends Parasitol 18:: 405--410.
    • (2002) Trends Parasitol , vol.18 , pp. 405-410
    • Tibayrenc, M.1    Ayala, F.J.2
  • 6
    • 0017854251 scopus 로고
    • Isozymic heterogeneity of Trypanosoma cruzi in the first autochthonous patients with Chagas' disease in Amazonian Brazil
    • Miles MA, Souza A, Povoa M, Shaw JJ, Lainson R, et al. ((1978)) Isozymic heterogeneity of Trypanosoma cruzi in the first autochthonous patients with Chagas' disease in Amazonian Brazil. Nature 272:: 819--821.
    • (1978) Nature , vol.272 , pp. 819-821
    • Miles, M.A.1    Souza, A.2    Povoa, M.3    Shaw, J.J.4    Lainson, R.5
  • 8
    • 0034603698 scopus 로고    scopus 로고
    • Evolutionary relationships in Trypanosoma cruzi: molecular phylogenetics supports the existence of a new major lineage of strains
    • Robello C, Gamarro F, Castanys S, Alvarez Valin F, ((2000)) Evolutionary relationships in Trypanosoma cruzi: molecular phylogenetics supports the existence of a new major lineage of strains. Gene 246:: 331--338.
    • (2000) Gene , vol.246 , pp. 331-338
    • Robello, C.1    Gamarro, F.2    Castanys, S.3    Alvarez Valin, F.4
  • 10
    • 0034071224 scopus 로고    scopus 로고
    • Population structure and genetic typing of Trypanosoma cruzi, the agent of Chagas disease: a multilocus enzyme electrophoresis approach
    • Barnabé C, Brisse S, Tibayrenc M, ((2000)) Population structure and genetic typing of Trypanosoma cruzi, the agent of Chagas disease: a multilocus enzyme electrophoresis approach. Parasitology 120 (Pt 5): 513--526.
    • (2000) Parasitology , vol.120 , Issue.Pt 5 , pp. 513-526
    • Barnabé, C.1    Brisse, S.2    Tibayrenc, M.3
  • 11
    • 0033954610 scopus 로고    scopus 로고
    • Identification of six Trypanosoma cruzi phylogenetic lineages by random amplified polymorphic DNA and multilocus enzyme electrophoresis
    • Brisse S, Barnabé C, Tibayrenc M, ((2000)) Identification of six Trypanosoma cruzi phylogenetic lineages by random amplified polymorphic DNA and multilocus enzyme electrophoresis. Int J Parasitol 30:: 35--44.
    • (2000) Int J Parasitol , vol.30 , pp. 35-44
    • Brisse, S.1    Barnabé, C.2    Tibayrenc, M.3
  • 12
    • 0034881536 scopus 로고    scopus 로고
    • Characterisation of large and small subunit rRNA and mini-exon genes further supports the distinction of six Trypanosoma cruzi lineages
    • Brisse S, Verhoef J, Tibayrenc M, ((2001)) Characterisation of large and small subunit rRNA and mini-exon genes further supports the distinction of six Trypanosoma cruzi lineages. Int J Parasitol 31:: 1218--1226.
    • (2001) Int J Parasitol , vol.31 , pp. 1218-1226
    • Brisse, S.1    Verhoef, J.2    Tibayrenc, M.3
  • 13
    • 71149119832 scopus 로고    scopus 로고
    • A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI
    • Zingales B, Andrade SG, Briones MRS, Campbell DA, Chiari E, et al. ((2009)) A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI. Mem Inst Oswaldo Cruz 104:: 1051--1054.
    • (2009) Mem Inst Oswaldo Cruz , vol.104 , pp. 1051-1054
    • Zingales, B.1    Andrade, S.G.2    Briones, M.R.S.3    Campbell, D.A.4    Chiari, E.5
  • 14
    • 84857655347 scopus 로고    scopus 로고
    • The revised Trypanosoma cruzi subspecific nomenclature: rationale, epidemiological relevance and research applications
    • Zingales B, Miles MA, Campbell DA, Tibayrenc M, Macedo AM, et al. ((2012)) The revised Trypanosoma cruzi subspecific nomenclature: rationale, epidemiological relevance and research applications. Infect Genet Evol 12:: 240--253.
    • (2012) Infect Genet Evol , vol.12 , pp. 240-253
    • Zingales, B.1    Miles, M.A.2    Campbell, D.A.3    Tibayrenc, M.4    Macedo, A.M.5
  • 15
    • 67651154550 scopus 로고    scopus 로고
    • A new genotype of Trypanosoma cruzi associated with bats evidenced by phylogenetic analyses using SSU rDNA, cytochrome b and Histone H2B genes and genotyping based on ITS1 rDNA
    • Marcili A, Lima L, Cavazzana M, Junqueira ACV, Veludo HH, et al. ((2009)) A new genotype of Trypanosoma cruzi associated with bats evidenced by phylogenetic analyses using SSU rDNA, cytochrome b and Histone H2B genes and genotyping based on ITS1 rDNA. Parasitology 136:: 641--655.
    • (2009) Parasitology , vol.136 , pp. 641-655
    • Marcili, A.1    Lima, L.2    Cavazzana, M.3    Junqueira, A.C.V.4    Veludo, H.H.5
  • 16
    • 79959858450 scopus 로고    scopus 로고
    • Multilocus sequence typing (MLST) for lineage assignment and high resolution diversity studies in Trypanosoma cruzi
    • Yeo M, Mauricio IL, Messenger LA, Lewis MD, Llewellyn MS, et al. ((2011)) Multilocus sequence typing (MLST) for lineage assignment and high resolution diversity studies in Trypanosoma cruzi. PLoS Negl Trop Dis 5:: e1049.
    • (2011) PLoS Negl Trop Dis , vol.5
    • Yeo, M.1    Mauricio, I.L.2    Messenger, L.A.3    Lewis, M.D.4    Llewellyn, M.S.5
  • 17
    • 84855998981 scopus 로고    scopus 로고
    • Analyses of 32 loci clarify phylogenetic relationships among Trypanosoma cruzi lineages and support a single hybridization prior to human contact
    • Flores López CA, Machado CA, ((2011)) Analyses of 32 loci clarify phylogenetic relationships among Trypanosoma cruzi lineages and support a single hybridization prior to human contact. PLoS Negl Trop Dis 5:: e1272.
    • (2011) PLoS Negl Trop Dis , vol.5
    • Flores López, C.A.1    Machado, C.A.2
  • 18
    • 0035912819 scopus 로고    scopus 로고
    • Nucleotide sequences provide evidence of genetic exchange among distantly related lineages of Trypanosoma cruzi
    • Machado CA, Ayala FJ, ((2001)) Nucleotide sequences provide evidence of genetic exchange among distantly related lineages of Trypanosoma cruzi. Proc Natl Acad Sci U S A 98:: 7396--7401.
    • (2001) Proc Natl Acad Sci U S A , vol.98 , pp. 7396-7401
    • Machado, C.A.1    Ayala, F.J.2
  • 19
    • 31544467265 scopus 로고    scopus 로고
    • Two hybridization events define the population structure of Trypanosoma cruzi
    • Westenberger SJ, Barnabé C, Campbell DA, Sturm NR, ((2005)) Two hybridization events define the population structure of Trypanosoma cruzi. Genetics 171:: 527--543.
    • (2005) Genetics , vol.171 , pp. 527-543
    • Westenberger, S.J.1    Barnabé, C.2    Campbell, D.A.3    Sturm, N.R.4
  • 20
    • 0020988467 scopus 로고
    • Correlation between isoenzyme patterns and biological behaviour of different strains of Trypanosoma cruzi
    • Andrade V, Brodskyn C, Andrade SG, ((1983)) Correlation between isoenzyme patterns and biological behaviour of different strains of Trypanosoma cruzi. Trans R Soc Trop Med Hyg 77:: 796--799.
    • (1983) Trans R Soc Trop Med Hyg , vol.77 , pp. 796-799
    • Andrade, V.1    Brodskyn, C.2    Andrade, S.G.3
  • 21
    • 0032078462 scopus 로고    scopus 로고
    • Trypanosoma cruzi: impact of clonal evolution of the parasite on its biological and medical properties
    • Revollo S, Oury B, Laurent JP, Barnabé C, Quesney V, et al. ((1998)) Trypanosoma cruzi: impact of clonal evolution of the parasite on its biological and medical properties. Exp Parasitol 89:: 30--39.
    • (1998) Exp Parasitol , vol.89 , pp. 30-39
    • Revollo, S.1    Oury, B.2    Laurent, J.P.3    Barnabé, C.4    Quesney, V.5
  • 22
    • 0019423055 scopus 로고
    • Do radically dissimilar Trypanosoma cruzi strains (zymodemes) cause Venezuelan and Brazilian forms of Chagas' disease
    • Miles MA, Cedillos RA, Póvoa MM, de Souza AA, Prata A, et al. ((1981)) Do radically dissimilar Trypanosoma cruzi strains (zymodemes) cause Venezuelan and Brazilian forms of Chagas' disease? Lancet 1:: 1338--1340.
    • (1981) Lancet , vol.1 , pp. 1338-1340
    • Miles, M.A.1    Cedillos, R.A.2    Póvoa, M.M.3    de Souza, A.A.4    Prata, A.5
  • 23
    • 0032032285 scopus 로고    scopus 로고
    • Genetic Variability of Trypanosoma cruzi:Implications for the Pathogenesis of Chagas Disease
    • Macedo AM, Pena SD, ((1998)) Genetic Variability of Trypanosoma cruzi:Implications for the Pathogenesis of Chagas Disease. Parasitol Today 14:: 119--124.
    • (1998) Parasitol Today , vol.14 , pp. 119-124
    • Macedo, A.M.1    Pena, S.D.2
  • 24
    • 0032940097 scopus 로고    scopus 로고
    • Differential tissue distribution of diverse clones of Trypanosoma cruzi in infected mice
    • Andrade LO, Machado CR, Chiari E, Pena SD, Macedo AM, ((1999)) Differential tissue distribution of diverse clones of Trypanosoma cruzi in infected mice. Mol Biochem Parasitol 100:: 163--172.
    • (1999) Mol Biochem Parasitol , vol.100 , pp. 163-172
    • Andrade, L.O.1    Machado, C.R.2    Chiari, E.3    Pena, S.D.4    Macedo, A.M.5
  • 25
    • 0033835153 scopus 로고    scopus 로고
    • Genetic characterization of Trypanosoma cruzi directly from tissues of patients with chronic Chagas disease: differential distribution of genetic types into diverse organs
    • Vago AR, Andrade LO, Leite AA, d'Avila Reis D, Macedo AM, et al. ((2000)) Genetic characterization of Trypanosoma cruzi directly from tissues of patients with chronic Chagas disease: differential distribution of genetic types into diverse organs. Am J Pathol 156:: 1805--1809.
    • (2000) Am J Pathol , vol.156 , pp. 1805-1809
    • Vago, A.R.1    Andrade, L.O.2    Leite, A.A.3    d'Avila Reis, D.4    Macedo, A.M.5
  • 26
    • 73949161043 scopus 로고    scopus 로고
    • Genotyping of Trypanosoma cruzi: systematic selection of assays allowing rapid and accurate discrimination of all known lineages
    • Lewis MD, Ma J, Yeo M, Carrasco HJ, Llewellyn MS, et al. ((2009)) Genotyping of Trypanosoma cruzi: systematic selection of assays allowing rapid and accurate discrimination of all known lineages. Am J Trop Med Hyg 81:: 1041--1049.
    • (2009) Am J Trop Med Hyg , vol.81 , pp. 1041-1049
    • Lewis, M.D.1    Ma, J.2    Yeo, M.3    Carrasco, H.J.4    Llewellyn, M.S.5
  • 27
    • 79956319236 scopus 로고    scopus 로고
    • Further interest of miniexon multiplex PCR for a rapid typing of Trypanosoma cruzi DTU groups
    • Aliaga C, Brenière SF, Barnabé C, ((2011)) Further interest of miniexon multiplex PCR for a rapid typing of Trypanosoma cruzi DTU groups. Infect Genet Evol 11:: 1155--1158.
    • (2011) Infect Genet Evol , vol.11 , pp. 1155-1158
    • Aliaga, C.1    Brenière, S.F.2    Barnabé, C.3
  • 28
    • 0027497964 scopus 로고
    • Genetic characterization of six parasitic protozoa: parity between random-primer DNA typing and multilocus enzyme electrophoresis
    • Tibayrenc M, Neubauer K, Barnabé C, Guerrini F, Skarecky D, et al. ((1993)) Genetic characterization of six parasitic protozoa: parity between random-primer DNA typing and multilocus enzyme electrophoresis. Proc Natl Acad Sci U S A 90:: 1335--1339.
    • (1993) Proc Natl Acad Sci U S A , vol.90 , pp. 1335-1339
    • Tibayrenc, M.1    Neubauer, K.2    Barnabé, C.3    Guerrini, F.4    Skarecky, D.5
  • 29
    • 84857644998 scopus 로고    scopus 로고
    • Candidate targets for Multilocus Sequence Typing of Trypanosoma cruzi: Validation using parasite stocks from the Chaco Region and a set of reference strains
    • Lauthier JL, Tomasini N, Barnabé C, Monje Rumi MM, Alberti D'Amato AM, et al. ((2012)) Candidate targets for Multilocus Sequence Typing of Trypanosoma cruzi: Validation using parasite stocks from the Chaco Region and a set of reference strains. Infect Genet Evol 12:: 350--358.
    • (2012) Infect Genet Evol , vol.12 , pp. 350-358
    • Lauthier, J.L.1    Tomasini, N.2    Barnabé, C.3    Monje Rumi, M.M.4    Alberti D'Amato, A.M.5
  • 30
    • 84859003689 scopus 로고    scopus 로고
    • Evolutionary Insights from Bat Trypanosomes: Morphological, Developmental and Phylogenetic Evidence of a New Species, Trypanosoma (Schizotrypanum) erneyi sp. nov., in African Bats Closely Related to Trypanosoma (Schizotrypanum) cruzi and Allied Species
    • Lima L, da Silva FM, Neves L, Attias M, Takata CSA, et al. ((2012)) Evolutionary Insights from Bat Trypanosomes: Morphological, Developmental and Phylogenetic Evidence of a New Species, Trypanosoma (Schizotrypanum) erneyi sp. nov., in African Bats Closely Related to Trypanosoma (Schizotrypanum) cruzi and Allied Species. Protist in press.
    • (2012) Protist
    • Lima, L.1    da Silva, F.M.2    Neves, L.3    Attias, M.4    Takata, C.S.A.5
  • 31
    • 0031955518 scopus 로고    scopus 로고
    • Base-calling of automated sequencer traces using phred. I. Accuracy assessment
    • Ewing B, Hillier L, Wendl MC, Green P, ((1998)) Base-calling of automated sequencer traces using phred. I. Accuracy assessment. Genome Res 8:: 175--185.
    • (1998) Genome Res , vol.8 , pp. 175-185
    • Ewing, B.1    Hillier, L.2    Wendl, M.C.3    Green, P.4
  • 32
    • 0031978181 scopus 로고    scopus 로고
    • Base-calling of automated sequencer traces using phred. II. Error probabilities
    • Ewing B, Green P, ((1998)) Base-calling of automated sequencer traces using phred. II. Error probabilities. Genome Res 8:: 186--194.
    • (1998) Genome Res , vol.8 , pp. 186-194
    • Ewing, B.1    Green, P.2
  • 33
    • 0030872838 scopus 로고    scopus 로고
    • PolyPhred: automating the detection and genotyping of single nucleotide substitutions using uorescence-based resequencing
    • Nickerson DA, Tobe VO, Taylor SL, ((1997)) PolyPhred: automating the detection and genotyping of single nucleotide substitutions using uorescence-based resequencing. Nucleic Acids Res 25:: 2745--2751.
    • (1997) Nucleic Acids Res , vol.25 , pp. 2745-2751
    • Nickerson, D.A.1    Tobe, V.O.2    Taylor, S.L.3
  • 34
    • 43349108713 scopus 로고    scopus 로고
    • Viewing and editing assembled sequences using Consed
    • Unit11
    • Gordon D, ((2003)) Viewing and editing assembled sequences using Consed. Curr Protoc Bioinformatics Chapter 11:: Unit11.2.
    • (2003) Curr Protoc Bioinformatics Chapter , vol.11 , pp. 2
    • Gordon, D.1
  • 35
    • 58149204065 scopus 로고    scopus 로고
    • TcSNP: a database of genetic variation in Trypanosoma cruzi
    • Ackermann AA, Carmona SJ, Agüero F, ((2009)) TcSNP: a database of genetic variation in Trypanosoma cruzi. Nucleic Acids Res 37:: D544--D549.
    • (2009) Nucleic Acids Res , vol.37
    • Ackermann, A.A.1    Carmona, S.J.2    Agüero, F.3
  • 36
    • 67649401919 scopus 로고    scopus 로고
    • Chromosome level assembly of the hybrid Trypanosoma cruzi genome
    • Weatherly BD, Boehlke C, Tarleton RL, ((2009)) Chromosome level assembly of the hybrid Trypanosoma cruzi genome. BMC Genomics 10:: 255.
    • (2009) BMC Genomics , vol.10 , pp. 255
    • Weatherly, B.D.1    Boehlke, C.2    Tarleton, R.L.3
  • 38
    • 0027745825 scopus 로고
    • Effect of PCR conditions on the formation of heteroduplex and single-stranded DNA products in the amplification of bacterial ribosomal DNA spacer regions
    • Jensen MA, Straus N, ((1993)) Effect of PCR conditions on the formation of heteroduplex and single-stranded DNA products in the amplification of bacterial ribosomal DNA spacer regions. PCR Methods Appl 3:: 186--194.
    • (1993) PCR Methods Appl , vol.3 , pp. 186-194
    • Jensen, M.A.1    Straus, N.2
  • 39
    • 0029972052 scopus 로고    scopus 로고
    • The frequency of chimeric molecules as a consequence of PCR coamplification of 16S rRNA genes from different bacterial species
    • Wang GC, Wang Y, ((1996)) The frequency of chimeric molecules as a consequence of PCR coamplification of 16S rRNA genes from different bacterial species. Microbiology 142 (Pt 5): 1107--1114.
    • (1996) Microbiology , vol.142 , Issue.Pt 5 , pp. 1107-1114
    • Wang, G.C.1    Wang, Y.2
  • 40
    • 0036566806 scopus 로고    scopus 로고
    • Heteroduplexes in mixed-template amplifications: formation, consequence and elimination by 'reconditioning PCR
    • Thompson JR, Marcelino LA, Polz MF, ((2002)) Heteroduplexes in mixed-template amplifications: formation, consequence and elimination by 'reconditioning PCR'. Nucleic Acids Res 30:: 2083--2088.
    • (2002) Nucleic Acids Res , vol.30 , pp. 2083-2088
    • Thompson, J.R.1    Marcelino, L.A.2    Polz, M.F.3
  • 41
    • 34247539877 scopus 로고    scopus 로고
    • Multilocus polymerase chain reaction restriction fragment-length polymorphism genotyping of Trypanosoma cruzi (Chagas disease): taxonomic and clinical applications
    • Rozas M, De Doncker S, Adaui V, Coronado X, Barnabé C, et al. ((2007)) Multilocus polymerase chain reaction restriction fragment-length polymorphism genotyping of Trypanosoma cruzi (Chagas disease): taxonomic and clinical applications. J Infect Dis 195:: 1381--1388.
    • (2007) J Infect Dis , vol.195 , pp. 1381-1388
    • Rozas, M.1    de Doncker, S.2    Adaui, V.3    Coronado, X.4    Barnabé, C.5
  • 42
    • 0008894099 scopus 로고
    • Strains and clones of Trypanosoma cruzi can be characterized by pattern of restriction endonuclease products of kinetoplast DNA minicircles
    • Morel C, Chiari E, Camargo EP, Mattei DM, Romanha AJ, et al. ((1980)) Strains and clones of Trypanosoma cruzi can be characterized by pattern of restriction endonuclease products of kinetoplast DNA minicircles. Proc Natl Acad Sci U S A 77:: 6810--6814.
    • (1980) Proc Natl Acad Sci U S A , vol.77 , pp. 6810-6814
    • Morel, C.1    Chiari, E.2    Camargo, E.P.3    Mattei, D.M.4    Romanha, A.J.5
  • 43
    • 34548014064 scopus 로고    scopus 로고
    • Direct molecular profiling of minicircle signatures and lineages of Trypanosoma cruzi bloodstream populations causing congenital Chagas disease
    • Burgos JM, Altcheh J, Bisio M, Duffy T, Valadares HMS, et al. ((2007)) Direct molecular profiling of minicircle signatures and lineages of Trypanosoma cruzi bloodstream populations causing congenital Chagas disease. Int J Parasitol 37:: 1319--1327.
    • (2007) Int J Parasitol , vol.37 , pp. 1319-1327
    • Burgos, J.M.1    Altcheh, J.2    Bisio, M.3    Duffy, T.4    Valadares, H.M.S.5
  • 44
    • 66749136414 scopus 로고    scopus 로고
    • Probing population dynamics of Trypanosoma cruzi during progression of the chronic phase in chagasic patients
    • Alchaar D'Avila D, Macedo AM, Silva Valadares HM, Dias Gontijo E, de Castro AM, et al. ((2009)) Probing population dynamics of Trypanosoma cruzi during progression of the chronic phase in chagasic patients. J Clin Microbiol 47:: 1718--1725.
    • (2009) J Clin Microbiol , vol.47 , pp. 1718-1725
    • Alchaar D'Avila, D.1    Macedo, A.M.2    Silva Valadares, H.M.3    Dias Gontijo, E.4    de Castro, A.M.5
  • 45
    • 80051677176 scopus 로고    scopus 로고
    • Molecular diversity of the Trypanosoma cruzi TcSMUG family of mucin genes and proteins
    • Urban I, Santurio LB, Chidichimo A, Yu H, Chen X, et al. ((2011)) Molecular diversity of the Trypanosoma cruzi TcSMUG family of mucin genes and proteins. Biochem J 438:: 303--313.
    • (2011) Biochem J , vol.438 , pp. 303-313
    • Urban, I.1    Santurio, L.B.2    Chidichimo, A.3    Yu, H.4    Chen, X.5


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.