Molecular Biological Characteristics of Vibrio cholerae O1 Isolated from Diarrheal patients in the Gyeongbuk province.

최근 경북지역 설사환자 검체에서 분리된 Vibrio cholerae O1의 분자생물학적 특성

  • 이상조 (경북 보건환경연구원 미생물과) ;
  • 이복권 (국립보건원 세균질환부 장내세균과) ;
  • 이건주 (안동대학교 생물학과) ;
  • 이희무 (안동대학교 생물학과)
  • Published : 2003.12.01

Abstract

This study was carried out to investigate the cause of cholera outbreak in Gyeongbuk province in 2001.90 strains of Vibrio cholerae O1 El Tor serotype Inaba were isolated from diarrheal patients. By multiplex-PCR, all of the isolated strains revealed positive for detection ctxA, hlyA and tcpA genes. There were DNA sequence difference of the cholera-toxin subunit A gene and subunit B gene between isolated V. cholerae O1 and the strain of GenBank. In analysis of PFGE patterns, all of the isolated strains were showed the same DNA fragments. We also collected plankton samples in the east coast of Gyeongbuk to isolate V. cholerae O1 and V. cholerae O139 from August to October 2002. The samples were examined to detect the rfb gene and cholera-toxin gene by multiplex-PCR. The cholera-toxin gene was detected and then we tried to isolate V. cholerae O1 and V. cholerae O139, but they were not isolated.

2001년 경북지역 콜레라 유행시 설사환자에서 V. cholerae O1 El Tor형 90주를 분리하였다. 분리균을 대상으로 ctxA, tcpA, hlyA gene에 대한 multiplex-PCR 시험에서는 분리균 모두에서 302, 223, 471 bps크기의 DNA 증폭산물 band를 확인하였다. ctxA 및 ctxB gene 부위의 유전자 염기서열 비교에서 분리균과 GenBank에 수록된 균간의 유형의 차이를 보였다. PFGE typing 실험에서는 분리균 모두에서 동일한 amplicon 단편을 나타내었다. 또한, 2002년 8월부터 10월까지 콜레라균 서식가능성 규명을 위해 경북 동해안 일대에서 plankton 시료채취 및 V. cholerae O1 및 V. cholerae O139의 rfb 및 CT gene 검출을 위한 multiplex-PCR 확인실험에서는 ctxA gene의 DNA band는 일부 검출되었으나, V. cholerae O1 및 V. cholerae O139는 분리되지 않았다. 험에서 ctxA gene DNA band는 검출되었으나, V. cholerae O1 및 V. cholerae O1는 분리되지 않았다.

Keywords

References

  1. Manual of clinical microbiology(6th ed.) Murray,P.R.;Baron,E.J.;Michael,A.;Tenover,F.C.;Yolken,R.H.
  2. Vibrio manual of clinical microbiology (4th Ed.) Farmer,J.;J.Michael;T.Kelly;F.W.Hickman-Brenner
  3. 안동대 대학원 박사학위논문 국내에서 분리된 V.cholerae O1과 O139균의 분자역학적 특성 이복권
  4. 대한감염학회지 v.28 no.6 최근 국내 유입 V.cholerae균 및 1995년도 국내 집단 발생 콜레라의 역학적 양상 김호훈;신영학;강연호;유천권;박미선;김동숙;유재연;전정훈;이복권;박기덕;김동진;정태화;이종구;박기동;김상순;이동모;김문식;조병률
  5. 대한미생물학회지 v.27 no.4 1991년 한국의 콜레라 유행기간중 분리된 V. cholerae O1의 생화학적성상 및 장독소 유전자에 관하여 유천권;강윤숙;조수열;이영희;김기상;이명원;김호훈;박기덕
  6. 국립보건원보 v.18 Phage type of Vibrio cholerae cultures isolated in Korea(1980) 이복권;강연숙;정태화
  7. Bull. Wld. Hlth Org. v.28 Study on the haemolutic activity of El Tor Vibrio Feeley,J.C.;Margaret,P.
  8. 1991년도 보건학종합학술대회지 우리나라 콜레라 유행의 역학적 고찰 김정순
  9. 국립보건원보 v.3 세균검사사업-1963 년도 콜레라 방역보고 이종승;박기덕
  10. 한국역학회지 v.2 no.1 콜레라의 자연사 정희영
  11. 한국역학회지 v.13 no.2 1991년. 서천, 군산지역 콜레라 유행의 전파경로와 발생근원 김한중;오희철;서일;박종구;김규상;지선하;문종국
  12. 감염병 발생정보 v.12 no.10 2001년 발생한 콜레라 유행의 역학적 특성-전국 국립보건원
  13. Bio. Technology v.3 Viable, but non-culturable Vibrio cholerae and related pathgens in the environment: implication for release of genetically engineered microorganisms Colwell,R.R.;P.R.Braytom;D.J.Grimes;D.R.Roszak;S.A.Huq;L.M.Palmer https://doi.org/10.1038/nbt0985-817
  14. 한국감염학회 v.18 no.2 우리나라 1995, 1996년 콜레라 발생 근원에 대한 연구 오희철;김문식;이종구;김상순;박기동;김호훈;황창용
  15. Appl. Environ. Microbiol. v.45 no.1 Ecological relationship between V. cholerae and planktonic crustcean copepods Huq,A.;E.B.Small;P.A.West;M.I.Huq;R.Rahman;E.E.Clowell
  16. Appl. Environ. Microbiol v.48 no.2 Inflience of water temperature salinity, and pH on survival and growth of toxigenic V.cholerae serovar O1 associated with live copepods in laboratory microcosm Huq,A.;P.A.West;E.B.Small;M.I.Huq;R.R.Colwell
  17. Appl. Environ. Microbiol v.56 no.6 Attachment of V.cholerae serogroup O1 to zooplankton and phytolpankton of Bangladesh waters Tamplin,M.L.;A.L.Gauzens;A.Huq;D.A.Sack;R.R.Colwell
  18. Appl. Environ. Microbiol. v.50 no.2 Incidence of V. cholerae and relted Vibrios in coactal Lagoon and sea water influenced by lake discharges along an annual cycle Garay,E.;A.Arnau;C.Amaro
  19. Appl. Environ. Microbiol v.55 no.1 Isolation of Vibrio cholerae from aquatic birds in Colorado and Utah Ogg,J.E.;R.A.Ryder;H.L.Smith,Jr.
  20. Microb. Sci. v.3 The fate of enteric bacteria in estuarine and marine environment Grimes,D.J.;R.A.Atwell;P.R.Brayton;L.M.Palmer;D.M.Rollins;D.B.Roszak;F.L.Singleton;M.L.Tamplin,R.R.Colwell
  21. Appl. Environ. Microbiol. v.41 Isolation of Vibrio cholerae O1 from the eastern oyster Crassostrea viginea Hood,M.A.;G.E.Ness;G.E.Roderick
  22. Lancet v.326 Cholera epidemiology in developed and developing countries:new thoughts on transmission, seasonality, and control Miller,C.J.
  23. Cad Saude Publica v.12 no.4 Influence of the copepod Mesocyclops longisetus (Crustacea: Cyclopidae) on the survival of Vibrio cholerae O1 in fresh water Araujo,D.B.;S.C.Martins;L.M.Albuquerque;E.Hofer https://doi.org/10.1590/S0102-311X1996000400015
  24. Appl. Environ. Microbiol v.63 no.8 Effect of alum on free-living and copepod-associated Vibrio cholerae O1 and O139 Chowdhury,M.A.;A.Huq;B.Xu;F.J.Madeira;R.R.Colwell
  25. Comp. Immunol. Microbiol. Infect. Dis. v.19 no.3 Ecological relationship between Aeromonas and Vibrio spp. and planktonic copepods in the coastal marine environment in southern Italy Durnontet,S.;K.B.Krovacek;S.B.Aloda;R.Grottoli;V.Pasquale;S.Vanucci https://doi.org/10.1016/0147-9571(96)00012-4
  26. 2002년도 해양환경 콜레라 감시 국립보건원
  27. Diverse, Non-epidemic serogroup strains of Vibrio cholerae Genetic analysis of the virulence regions, CTX f prophage and Vivrio pathogenicity island (VPI) Li,M.;Y.Chen;M.Kotetishvili;O.C.Stine;J.G.Jr.Morris;A.Sulakvelidze;S.Sozhamannan
  28. Department of Bacterial. Infections v.31 no.1 Comparison of cholera toxin genes(ctxAB) of non-O1 Vibrio cholerae strains 854(O139-bengal) and S7(O37) from two outbreaks Yamamoto,K.;V.G.Do;M.Xu;T.Ioda;T.Miwatani;M.J.Albert;T.Honda
  29. Biochem. Biophys, Acta. v.1090 no.1 Nucleotide sequence analysis of the CT operon of the Vibrio cholerae classical strain 569 B. Dams;E.M.De Wolf;W.Dierick https://doi.org/10.1016/0167-4781(91)90050-V
  30. Institute of pidemilogy and Microbiology, Chinese Academy of Preventive Medicine, P.O.Box5 ctxAB gene of strains of Vibrio chlerae isolated from China Rui,Y.;B.Kan;S.Gao
  31. The Insititute for Genomic Research v.9712 DNA sequence of both chromosomes of the cholera pathogen Vibrido cholerae Heidelberg,J.F.;J.A.Eisen;W.C.Nelson;O.White
  32. J. Misainmurhag Hoiji v.35 no.3 Cloning and nucleotide sequence analysis of the virulence gene. cassette from Vibrio cholerae KNIH002 isolated in Korea Shin,H.J.;Y.C.Park;Y.C.Kim
  33. J. Ckin. Microbiol. v.35 Rapid PFGE protocol for typing of E. coli O157:H7 and other gram-negative organism in 1 day Gautom,R.
  34. J. Med. Microbiol v.50 no.3 Molecular charcaterisation of rough variants of Vibrio chlerae isolated from hospitalised patients with diarrhoea Mitra,R.K.;R.K.Nandy;T.Ramamurthy;S.K.Bhattacharys;S.Yamasaki;T.Shimade;Y.Takeda;G.B.Nair
  35. J. Clin. Microbiol. v.30 no.8 Use of polymerase chain reaction for detection of toxigenic Vibrio cholerae O1 stain from the latin american cholera epidemic Fields,P.I.;P.Tanja;W.Kate;O.Orjan
  36. J. Clin. Microbiol v.35 no.6 Rapid detection Vibrio cholerae O139 bengal from stool specimens by PCR Albert,J.M.;I.Dilara;N.Shamsun;Q.Firdausi;F.Susanna;W.Andrej
  37. J. Clin. Microbiol v.29 Polymerase chain reaction for detection of cholera enterotoxin operon of Vibiro cholerae Shirai,H.;M.Nishibuchi;T.Ramamurthy;S.K.Bhattacharya;S.K.Pal;Y.Takada
  38. Vibrio cholerae and cholera-molecular to global prespectives Wachsmuth,K.;P.A.Blake;O.Olsvik
  39. J. Nature v.306 no.5943 Cholera toxin genes, nucleotide sequence, deletion analysis and vassine development Mekalanos,J.J.;D.J.Swartz;G.D.Pearson;N.Harford;F.Groyne;M. de Wild https://doi.org/10.1038/306551a0
  40. Infect. Immun. v.60 Cloning of a gene (zot) encoding a new toxin produceds by Vibrio cholerae Baudry,B.;A.Fasano;J.Ketlyand;J.B.Kaper
  41. J. Clin. Microbiol. v.31 Gene encoding zonula occludens toxin (zot) does not occur independently form cholera enterotoxin gene (ctx) in Vibiro cholerae Johnson,J.A.;J.G.Morris,Jr.;J.B.Kaper
  42. Proc. Natl. Acad. Sci v.90 Accessory cholerae enterotoxin(ace), the third toxin of a Vibiro cholerae virulence cassette Michele,T.;J.E.Galen;J.Michalski;A.Fasano;J.B.Kaper https://doi.org/10.1073/pnas.90.11.5267
  43. Lancet v.341 Detection and biotyping Vibrio cholerae O1 with Multiplex chain reaction Keasler,S.P.;R.H.Hall https://doi.org/10.1016/0140-6736(93)90792-F
  44. FEMS. Microbiol. Lett v.62 no.23 Production of monoclonal antibodies against a hernag-glutinin/protease of Vibrio cholerae Non-O1 Honda,T.;N.A.Hata;K.Lerpocasombat;T.Miwatani
  45. Infect. Immun. v.55 Enterotoxicity of EI Tor-like hemolysin of non-O1 Vibrio cholerae Ichinose,Y.;K.Yamamoto;N.Nakasone;M.J.Tanabe;T.Takeda;T.Miwatani;M.Iwanaga
  46. Description of a possible new extracellular virulence factor in nontoxigenic Vibrio cholerae O1 v.3 Spira,W.M.;D.A.Sack;P.J.Fedorka-Cray;S.Sanyal;J.Madden;B.McCardell
  47. 국립보건원보 v.36 플랑크톤 등 해양생물체 및 연안 해·하수에 대한 콜레라균 오염도 및 서식가능성 연구 이복권;강연호;박미선;유재연;김성한;김준영
  48. 대한감염학회·대한 화학요법학회 추계학술대회 콜레라 : 해양 토착문제 임현술
  49. N. Engl. J. Med v.302 Cholera-a possible endemic focus in the United States Blake,P.A.;R.E.Weaver;Hollis https://doi.org/10.1056/NEJM198002073020601
  50. Bulletin of the WHO v.58 The Queens Land cholera incident of 1977. 2. The epidermiological investigation Rogers,R.C.
  51. Appl. Environ. Microbiol v.45 no.1 Isolation of V.cholerae serotype Ogawa from Florida estuary Motes,M.L.;S.R.Zywno;A.DePaola;R.E.Becker;M.W.Presnell
  52. Control of Comminicable Disease in Man(15th ed.) Benenson,A.S.