Evaluation of an alkaline phosphatase-labeled DNA probe for enumeration of Vibrio vulnificus in Gulf Coast oysters

被引:32
作者
DePaola, A
Motes, ML
Cook, DW
Veazey, J
Garthright, WE
Blodgett, R
机构
[1] US FDA,RESIDENT POST,BATON ROUGE,LA
[2] US FDA,DIV MATH,WASHINGTON,DC 20204
关键词
DNA probe; enzyme immunoassay; oyster; Vibrio vulnificus;
D O I
10.1016/S0167-7012(97)00030-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A direct plating method and an FDA method were compared for enumeration of Vibrio vulnificus in Gulf Coast oysters. The direct plating method was based on hybridization of colony lifts and used an alkaline phosphatase-labeled oligonucleotide probe that targeted the cytolysin gene (Direct-VVAP). The FDA method employs most probable number analysis with confirmation of suspect isolates by enzyme immunoassay (MPN-EIA). Indigenous V. vulnificus levels in oysters harvested from Florida, Alabama and Louisiana throughout the year and in Gulf Coast market oysters ranged from <10 to 10(5) g(-1). Similar V. vulnificus levels (r=0.66) were found with these two procedures in freshly harvested oysters collected between April and October. Measurement variance was much greater, however, with the MPN procedure (0.118) than with the DNA probe procedure (0.004). In market oysters, the MPN-EIA procedure gave 0.4 log(10) higher estimates than the Direct-VVAP method, but the methods were closely correlated (r=0.83). Confirmation procedures were in agreement >90% of the time, as suspect isolates confirmed by one procedure were identified by the other procedure. Except when V. vulnificus densities are low (<10 g(-1)), the Direct-VVAP method provides an alternative procedure that is more rapid and precise than the MPN-EIA analysis. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:115 / 120
页数:6
相关论文
共 19 条
[1]  
*AM PUBL HLTH ASS, 1970, PROCEDURES EXAMINATI
[2]   EFFECTS OF HEATING, PH AND THERMORADIATION ON INACTIVATION OF VIBRIO-VULNIFICUS [J].
AMA, AA ;
HAMDY, MK ;
TOLEDO, RT .
FOOD MICROBIOLOGY, 1994, 11 (03) :215-227
[3]   DISEASE CAUSED BY A MARINE VIBRIO - CLINICAL CHARACTERISTICS AND EPIDEMIOLOGY [J].
BLAKE, PA ;
MERSON, MH ;
WEAVER, RE ;
HOLLIS, DG ;
HEUBLEIN, PC .
NEW ENGLAND JOURNAL OF MEDICINE, 1979, 300 (01) :1-5
[4]  
CARDARELLILEITE P, 1996, 96 ANN M AM SOC MICR, P51
[5]   ESTIMATION OF BACTERIAL DENSITIES BY MEANS OF THE MOST PROBABLE NUMBER [J].
COCHRAN, WG .
BIOMETRICS, 1950, 6 (02) :105-116
[6]   COLD-STORAGE AND MILD HEAT-TREATMENT AS PROCESSING AIDS TO REDUCE THE NUMBERS OF VIBRIO-VULNIFICUS IN RAW OYSTERS [J].
COOK, DW ;
RUPLE, AD .
JOURNAL OF FOOD PROTECTION, 1992, 55 (12) :985-989
[8]  
ELLIOT EL, 1992, FOOD DRUG ADM BACTER, P111
[9]  
HLADY G, 1994, P 1994 VIBR VULN WOR, P27
[10]  
HLADY GA, 1996, J INFECT DIS, V173, P1176