Identification and characterization of Clostridium perfringens using single target DNA microarray chip

被引：18

作者：

Al-Khaldi, SF

Villanueva, D

Chizhikov, V

机构：

[1] US FDA, Ctr Food Safety & Appl Nutr, Div Microbiol Studies, HSF 517, College Pk, MD 20740 USA

[2] US FDA, Ctr Biol Evaluat & Res, Rockville, MD 20857 USA

来源：

INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY | 2004年 / 91卷 / 03期

关键词：

Clostridium perfringens; single target DNA microarray chip; toxin genes;

D O I：

10.1016/j.ijfoodmicro.2003.07.009

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

A DNA microarray method was developed to identify the presence of toxin genes: encoding beta toxin (cpb), epsilon toxin (etx), enterotoxin (cpe), alpha toxin (cpa), and iota toxin (iA) in Clostridium perfringens. To build the DNA chip, each gene sequence was represented by one similar to 22-bp amino-modified oligonucleotide printed twice on aldehyde-coated slides. Multiplex PCR with Cy3 and Cy5-dCTP derivatized fluorescent nucleotides was used to label five genes and fluorescent probes were prepared. The PCR probes were denatured and single-strand-labeled DNAs were separated and purified using magnetic beads. The presence of toxin genes in C. perfringens was detected by hybridization of amplified ssDNA probes to oligonucleotides on the chip representing one target sequence of each toxin gene. The DNA chip was able to identify eight strains of C. perfringens. (C) 2003 Elsevier B.V. All rights reserved.

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收藏

页码：289 / 296

页数：8

相关论文

共 14 条

[1] 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

[2] Detection and genotyping of human group A rotaviruses by oligonucleotide microarray hybridization [J].

Chizhikov, V ;

Wagner, M ;

Ivshina, A ;

Hoshino, Y ;

Kapikian, AZ ;

Chumakov, K .

JOURNAL OF CLINICAL MICROBIOLOGY, 2002, 40 (07) :2398-2407

[3] Microarray analysis of microbial virulence factors [J].

Chizhikov, V ;

Rasooly, A ;

Chumakov, K ;

Levy, DD .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001, 67 (07) :3258-3263

[4] Evidence that the enterotoxin gene can be episomal in Clostridium perfringens isolates associated with non-food-borne human gastrointestinal diseases [J].

Collie, RE ;

McClane, BA .

JOURNAL OF CLINICAL MICROBIOLOGY, 1998, 36 (01) :30-36

[5] THE ENTEROTOXIN GENE (CPE) OF CLOSTRIDIUM-PERFRINGENS CAN BE CHROMOSOMAL OR PLASMID-BORNE [J].

CORNILLOT, E ;

SAINTJOANIS, B ;

DAUBE, G ;

KATAYAMA, S ;

GRANUM, PE ;

CANARD, B ;

COLE, ST .

MOLECULAR MICROBIOLOGY, 1995, 15 (04) :639-647

[6] ILEAL LOOP FLUID ACCUMULATION AND PRODUCTION OF DIARRHEA IN RABBITS BY CELL-FREE PRODUCTS OF CLOSTRIDIUM PERFRINGENS [J].

DUNCAN, CL ;

STRONG, DH .

JOURNAL OF BACTERIOLOGY, 1969, 100 (01) :86-&

[7] Molecular variation between the alpha-toxins from the type strain (NCTC 8237) and clinical isolates of Clostridium perfringens associated with disease in man and animals [J].

Ginter, A ;

Williamson, ED ;

Dessy, F ;

Coppe, P ;

Bullifent, H ;

Howells, A ;

Titball, RW .

MICROBIOLOGY-UK, 1996, 142 :191-198

[8] MOLECULAR-GENETIC ANALYSIS OF BETA-TOXIN OF CLOSTRIDIUM-PERFRINGENS REVEALS SEQUENCE HOMOLOGY WITH ALPHA-TOXIN, GAMMA-TOXIN, AND LEUKOCIDIN OF STAPHYLOCOCCUS-AUREUS [J].

HUNTER, SEC ;

BROWN, JE ;

OYSTON, PCF ;

SAKURAI, J ;

TITBALL, RW .

INFECTION AND IMMUNITY, 1993, 61 (09) :3958-3965

[9] PHOSPHOLIPASE-C AND HEMOLYTIC ACTIVITIES OF CLOSTRIDIUM-PERFRINGENS ALPHA-TOXIN CLONED IN ESCHERICHIA-COLI - SEQUENCE AND HOMOLOGY WITH A BACILLUS-CEREUS PHOSPHOLIPASE-C [J].

LESLIE, D ;

FAIRWEATHER, N ;

PICKARD, D ;

DOUGAN, G ;

KEHOE, M .

MOLECULAR MICROBIOLOGY, 1989, 3 (03) :383-392

[10]

MEER RR, 1997, AM J VET RES, V58, P705