Can the fliC PCR-restriction fragment length polymorphism technique replace classic serotyping methods for characterizing the H antigen of enterotoxigenic Escherichia coli strains?

被引:22
作者
Moreno, ACR
Guth, BEC
Martinez, MB [1 ]
机构
[1] Univ Sao Paulo, Sch Pharmaceut Sci, Sao Paulo, Brazil
[2] Univ Fed Sao Paulo, Dept Microbiol, Sao Paulo, Brazil
关键词
D O I
10.1128/JCM.44.4.1453-1458.2006
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
In this study, we performed fliC PCR-restriction fragment length polymorphism (RFLP) to investigate whether this technique would be better than classic serotyping for the characterization of the H antigen in enterotoxigenic Escherichia coli (ETEC) strains. We showed that the fliC genes from ETEC strains can be characterized by restriction analysis of their polymorphism. Only one allele of the fliC gene from ETEC strains was found for each flagellar antigen, with the exception of H21. Nonmotile strains could also be characterized using this molecular technique. Moreover, determination of the somatic antigen was guided by the identification of the flagellar antigen from previously unknown serotypes of ETEC strains by PCR-RFLP, thus reducing the number of anti-antigen 0 sera used. The PCR-RFLP technique proved to be faster than classic serotyping for the characterization of the E. coli H antigen, taking 2 days to complete instead of the 7 or more days using classic serotyping. In conclusion, the H molecular typing for Enterobacteriaceae members may become an important epidemiological tool for the characterization of the H antigen of E. coli pathotypes. The PCR-RFLP technique is capable of guiding the determination of the H antigen and could partially replace seroagglutination. With the determination of the molecular profiles of alleles from strains obtained in epidemiological studies, new patterns will be described for ETEC strains or other E. coli pathotypes, thus permitting widespread use of this technique to characterize fliC genes and determine the H antigen of E. coli strains.
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页码:1453 / 1458
页数:6
相关论文
共 18 条
[1]   Molecular typing and phylogenetic analysis of enteroinvasive Escherichia coli using the fliC gene sequence [J].
Amhaz, JMK ;
Andrade, A ;
Bando, SY ;
Tanaka, TL ;
Moreira-Filho, CA ;
Martinez, MB .
FEMS MICROBIOLOGY LETTERS, 2004, 235 (02) :259-264
[2]   Identification of EPEC and non-EPEC serotypes in the EPEC O serogroups by PCR-RFLP analysis of the fliC gene [J].
Botelho, BA ;
Bando, SY ;
Trabulsi, LR ;
Moreira, CA .
JOURNAL OF MICROBIOLOGICAL METHODS, 2003, 54 (01) :87-93
[3]   Clonal relationships among Shigella serotypes suggested by cryptic flagellin gene polymorphism [J].
Coimbra, RS ;
Lefevre, M ;
Grimont, F ;
Grimont, PAD .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (02) :670-674
[4]  
EWING WH, 1999, EDWARDS EWINGS IDENT
[5]   Molecular characterization of the gene encoding H antigen in Escherichia coli and development of a PCR-restriction fragment length polymorphism test for identification of E-coli O157:H7 and O157:NM [J].
Fields, PI ;
Blom, K ;
Hughes, HJ ;
Helsel, LO ;
Feng, P ;
Swaminathan, B .
JOURNAL OF CLINICAL MICROBIOLOGY, 1997, 35 (05) :1066-1070
[6]  
IINO T, 1977, ANNU REV GENET, V11, P161
[7]   THE FLAGELLAR FILAMENT PROTEIN [J].
JOYS, TM .
CANADIAN JOURNAL OF MICROBIOLOGY, 1988, 34 (04) :452-458
[8]   Molecular epidemiology of diarrhoeagenic Escherichia coli associated with sporadic cases and outbreaks of diarrhoea between 2000 and 2001 in India [J].
Kahali S. ;
Sarkar B. ;
Chakraborty S. ;
Macaden R. ;
Deokule J.S. ;
Ballal M. ;
Nandy R.K. ;
Bhattacharya S.K. ;
Takeda Y. ;
Ramamurthy T. .
European Journal of Epidemiology, 2004, 19 (5) :473-479
[9]   Identification of Escherichia coli flagellar types by restriction of the amplified fliC gene [J].
Machado, J ;
Grimont, F ;
Grimont, PAD .
RESEARCH IN MICROBIOLOGY, 2000, 151 (07) :535-546
[10]   Relationship between outer membrane protein and lipopolysaccharide profiles and serotypes of enterotoxigenic Escherichia coli isolated in Brazil [J].
Nishimura, LS ;
Ferreira, LCS ;
Pacheco, ABF ;
Guth, BEC .
FEMS MICROBIOLOGY LETTERS, 1996, 143 (2-3) :253-258