Rapid pulsed-field gel electrophoresis protocol for subtyping of Campylobacter jejuni

被引:300
作者
Ribot, EM [1 ]
Fitzgerald, C [1 ]
Kubota, K [1 ]
Swaminathan, B [1 ]
Barrett, TJ [1 ]
机构
[1] Ctr Dis Control & Prevent, Atlanta, GA 30333 USA
关键词
D O I
10.1128/JCM.39.5.1889-1894.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We developed a rapid pulsed field gel electrophoresis (PFGE) protocol for subtyping Campylobacter isolates based on the standardized protocols used by PulseNet laboratories for the subtyping of other food-borne bacterial pathogens. Various combinations of buffers, reagents, reaction conditions (e.g., cell suspension concentration, lysis time, lysis temperature, and restriction enzyme concentration), and electrophoretic parameters were evaluated in an effort to devise a protocol that is simple, rapid, and robust. PFGE analysis of Campylobacter isolates can be completed in 24 to 30 h using this protocol, whereas the most widely used current protocols require 3 to 4 days to complete. Comparison of PFGE patterns obtained in six laboratories showed that subtyping results obtained using this protocol are highly reproducible.
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收藏
页码:1889 / 1894
页数:6
相关论文
共 23 条
[1]   Campylobacter jejuni -: An emerging foodborne pathogen [J].
Altekruse, SF ;
Stern, NJ ;
Fields, PI ;
Swerdlow, DL .
EMERGING INFECTIOUS DISEASES, 1999, 5 (01) :28-35
[2]   DIFFERENTIATION OF CAMPYLOBACTER SPECIES USING PHENOTYPIC CHARACTERIZATION [J].
BARRETT, TJ ;
PATTON, CM ;
MORRIS, GK .
LABORATORY MEDICINE, 1988, 19 (02) :96-102
[3]  
BOLTON FJ, 1996, CAMPYLOBACTERS HELIC, P1
[4]  
*CDCP, 2000, QUAL ASS QA QUAL CON
[5]  
Centers for Disease Control and Prevention (CDC), 2000, STAND MOL SUBT FOODB
[6]   USE OF PULSED-FIELD AGAROSE-GEL ELECTROPHORESIS TO SIZE GENOMES OF CAMPYLOBACTER SPECIES AND TO CONSTRUCT A SALI MAP OF CAMPYLOBACTER-JEJUNI UA580 [J].
CHANG, N ;
TAYLOR, DE .
JOURNAL OF BACTERIOLOGY, 1990, 172 (09) :5211-5217
[7]   AN OUTBREAK OF CAMPYLOBACTER-JEJUNI ENTERITIS ASSOCIATED WITH FAILED MILK PASTEURIZATION [J].
FAHEY, T ;
MORGAN, D ;
GUNNEBURG, C ;
ADAK, GK ;
MAJID, F ;
KACZMARSKI, E .
JOURNAL OF INFECTION, 1995, 31 (02) :137-143
[8]   COMPARISON OF PFGE, RIBOTYPING AND PHAGE-TYPING IN THE EPIDEMIOLOGIC ANALYSIS OF CAMPYLOBACTER-JEJUNI SEROTYPE HS2 INFECTIONS [J].
GIBSON, JR ;
FITZGERALD, C ;
OWEN, RJ .
EPIDEMIOLOGY AND INFECTION, 1995, 115 (02) :215-225
[9]   MOLECULAR EPIDEMIOLOGY - APPLICATION OF CONTEMPORARY TECHNIQUES TO THE TYPING OF MICROORGANISMS [J].
MASLOW, JN ;
MULLIGAN, ME ;
ARBEIT, RD .
CLINICAL INFECTIOUS DISEASES, 1993, 17 (02) :153-164
[10]   Food-related illness and death in the United States [J].
Mead, PS ;
Slutsker, L ;
Dietz, V ;
McCaig, LF ;
Bresee, JS ;
Shapiro, C ;
Griffin, PM ;
Tauxe, RV .
EMERGING INFECTIOUS DISEASES, 1999, 5 (05) :607-625