Detection of Vibrio vulnificus biotypes 1 and 2 in eels and oysters by PCR amplification

被引:29
作者
Coleman, SS
Melanson, DM
Biosca, EG
Oliver, JD
机构
[1] UNIV N CAROLINA,DEPT BIOL,CHARLOTTE,NC 28223
[2] UNIV VALENCIA,DEPT MICROBIOL,E-46100 VALENCIA,SPAIN
关键词
D O I
10.1128/AEM.62.4.1378-1382.1996
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
DNA extraction procedures and PCR conditions to detect Vibrio vulnificus cells naturally occurring in oysters were developed. In addition, PCR amplification of V. vulnificus from oysters seeded with biotype 1 cells was demonstrated. By the methods described, V. vulnificus cells on a medium (colistin-polymyxin B-cellobiose agar) selective for this pathogen were detectable in oysters harvested in January and March, containing no culturable cells (<67 CFU/g), as well as in oysters harvested in May and June, containing culturable cells. It was possible to complete DNA extraction, PCR, and gel electrophoresis within 10 h by using the protocol described for oysters. V. vulnificus biotype 2 cells were also detected in eel tissues that had been infected with this strain and subsequently preserved in formalin. The protocol used for detection of V. vulnificus cells in eels required less than 5 h to complete. Optimum MgCl2 concentrations for the PCR of V. vulnificus from oysters and eels were different, although the same primer pair was used for both. This is the first report on the detection of cells of V. vulnificus naturally present in shellfish and represents a potentially powerful method for monitoring this important human and eel pathogen.
引用
收藏
页码:1378 / 1382
页数:5
相关论文
共 53 条
  • [11] POLYMERASE CHAIN REACTION-GENE PROBE DETECTION OF MICROORGANISMS BY USING FILTER-CONCENTRATED SAMPLES
    BEJ, AK
    MAHBUBANI, MH
    DICESARE, JL
    ATLAS, RM
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1991, 57 (12) : 3529 - 3534
  • [12] DETECTION OF COLIFORM BACTERIA IN WATER BY POLYMERASE CHAIN-REACTION AND GENE PROBES
    BEJ, AK
    STEFFAN, RJ
    DICESARE, J
    HAFF, L
    ATLAS, RM
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1990, 56 (02) : 307 - 314
  • [13] EVALUATION OF THE API 20E SYSTEM FOR IDENTIFICATION AND DISCRIMINATION OF VIBRIO-VULNIFICUS BIOTYPE-1 AND BIOTYPE-2
    BIOSCA, EG
    ESTEVE, C
    GARAY, E
    AMARO, C
    [J]. JOURNAL OF FISH DISEASES, 1993, 16 (01) : 79 - 82
  • [14] 1ST RECORD OF VIBRIO-VULNIFICUS BIOTYPE-2 FROM DISEASED EUROPEAN EEL, ANGUILLA-ANGUILLA L
    BIOSCA, EG
    AMARO, C
    ESTEVE, C
    ALCAIDE, E
    GARAY, E
    [J]. JOURNAL OF FISH DISEASES, 1991, 14 (01) : 103 - 109
  • [15] PRESENCE OF A CAPSULE IN VIBRIO-VULNIFICUS BIOTYPE-2 AND ITS RELATIONSHIP TO VIRULENCE FOR EELS
    BIOSCA, EG
    LLORENS, H
    GARAY, E
    AMARO, C
    [J]. INFECTION AND IMMUNITY, 1993, 61 (05) : 1611 - 1618
  • [16] BIOSCA EG, 1995, APPL ENVIRON MICROB, V62, P918
  • [17] DISEASE CAUSED BY A MARINE VIBRIO - CLINICAL CHARACTERISTICS AND EPIDEMIOLOGY
    BLAKE, PA
    MERSON, MH
    WEAVER, RE
    HOLLIS, DG
    HEUBLEIN, PC
    [J]. NEW ENGLAND JOURNAL OF MEDICINE, 1979, 300 (01) : 1 - 5
  • [18] USE OF THE POLYMERASE CHAIN-REACTION IN DETECTION OF CULTURABLE AND NONCULTURABLE VIBRIO-VULNIFICUS CELLS
    BRAUNS, LA
    HUDSON, MC
    OLIVER, JD
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1991, 57 (09) : 2651 - 2655
  • [19] BRAUNS LA, 1995, FOOD BIOTECHNOL, V8, P1
  • [20] Coleman SS, 1996, FEMS MICROBIOL ECOL, V19, P127, DOI 10.1016/0168-6496(95)00086-0