Development of a serotype-specific DNA Microarray for identification of some Shigella and pathogenic Escherichia coli strains

被引:48
作者
Li, Yayue
Liu, Dan
Cao, Boyang
Han, Weiqing
Liu, Yanqun
Liu, Fenxia
Guo, Xi
Bastin, David A.
Feng, Lu
Wang, Lei
机构
[1] Nankai Univ, TEDA Sch Biol Sci & Biotechnol, Tianjin 300457, Peoples R China
[2] Nankai Univ, TEDA Coll, Tianjin Res Ctr Funct Genom & Biochips, Tianjin 300457, Peoples R China
[3] Nankai Univ, TEDA Coll, Tianjin Key Lab Microbial Funct Genom, Tianjin 300457, Peoples R China
[4] Tianjin Biochip Corp, Tianjin 300457, Peoples R China
关键词
D O I
10.1128/JCM.01389-06
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Shigella and pathogenic Escherichia coli are major causes of human infectious diseases and are responsible for millions of cases of diarrhea worldwide every year. A convenient and rapid method to identify highly pathogenic serotypes of Shigella and E. coli is needed for large-scale epidemiologic study, timely clinical diagnosis, and reliable quarantine of the pathogens. In this study, a DNA microarray targeting O-serotype-specific genes was developed to detect 15 serotypes of Shigella and E. coli, including Shigella sonnei; Shigella flexneri type 2a; Shigella boydii types 7, 9, 13, 16, and 18; Shigella dysenteriae types 4, 8, and 10; and E. coli 055, O111, O114, O128, and O157. The microarray was tested against 186 representative strains of all Shigella and E. coli 0 serotypes, 38 clinical isolates, and 9 strains of other bacterial species that are commonly present in stool samples and was shown to be specific and reproducible. The detection sensitivity was 50 ng genomic DNA or 10(4) CFU per ml in mock stool specimens. This is the first report of a microarray for serotyping Shigella and pathogenic E. coli. The method has a number of advantages over traditional bacterial culture and antiserum agglutination methods and is promising for applications in basic microbiological research, clinical diagnosis, food safety, and epidemiological surveillance.
引用
收藏
页码:4376 / 4383
页数:8
相关论文
共 37 条
[1]   SEQUENCE AND ANALYSIS OF THE O-ANTIGEN GENE (RFB) CLUSTER OF ESCHERICHIA-COLI-O111 [J].
BASTIN, DA ;
REEVES, PR .
GENE, 1995, 164 (01) :17-23
[2]   Sequence analysis of the Escherichia coli O15 antigen gene cluster and development of a PCR assay for rapid detection of intestinal and extraintestinal pathogenic E. coli O15 strains [J].
Beutin, L ;
Tao, J ;
Feng, L ;
Krause, G ;
Zimmermann, S ;
Gleier, K ;
Xia, QY ;
Wang, L .
JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (02) :703-710
[3]   Development of PCR assays targeting the genes involved in synthesis and assembly of the new Escherichia coli O174 and O177O antigens [J].
Beutin, L ;
Kong, Q ;
Feng, L ;
Wang, Q ;
Krause, G ;
Leomil, L ;
Jin, Q ;
Wang, L .
JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (10) :5143-5149
[4]  
Bopp CA, 2003, MANUAL CLIN MICROBIO, P654
[5]   Detecting and genotyping Escherichia coli O157:H7 using multiplexed PCR and nucleic acid microarrays [J].
Call, DR ;
Brockman, FJ ;
Chandler, DP .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 2001, 67 (1-2) :71-80
[6]   Identification of Escherichia coli O-serogroups by restriction of the amplified O-antigen gene cluster (rfb-RFLP) [J].
Coimbra, RS ;
Grimont, F ;
Lenormand, P ;
Burguière, P ;
Beutin, L ;
Grimont, PAD .
RESEARCH IN MICROBIOLOGY, 2000, 151 (08) :639-654
[7]   Detection of Escherichia coli serogroups O26 and O113 by PCR amplification of the wzx and wzy genes [J].
DebRoy, C ;
Roberts, E ;
Kundrat, J ;
Davis, MA ;
Briggs, CE ;
Fratamico, PM .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (03) :1830-1832
[8]   Development of PCR assays targeting genes in O-antigen gene clusters for detection and identification of Escherichia coli O45 and O55 serogroups [J].
DebRoy, C ;
Fratamico, PM ;
Roberts, E ;
Davis, MA ;
Liu, YH .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (08) :4919-4924
[9]  
Ewing WH., 1986, EDWARDS EWINGS IDENT, V4th
[10]   Structure of the Shigella dysenteriae 7 O antigen gene cluster and identification of its antigen specific genes [J].
Feng, L ;
Tao, J ;
Guo, HJ ;
Xu, JG ;
Li, YY ;
Rezwan, F ;
Reeves, P ;
Wang, L .
MICROBIAL PATHOGENESIS, 2004, 36 (02) :109-115