Use of a DNA Microarray for Detection and Identification of Bacterial Pathogens Associated with Fishery Products

被引:21
作者
Cao, Boyang [1 ,2 ,3 ,4 ]
Li, Rongrong [1 ,2 ,3 ,4 ]
Xiong, Songjin [5 ]
Yao, Fangfang [1 ,2 ,3 ,4 ]
Liu, Xiangqian [1 ,2 ,3 ,4 ]
Wang, Min [1 ,2 ,3 ,4 ]
Feng, Lu [1 ,2 ,3 ,4 ]
Wang, Lei [1 ,2 ,3 ,4 ,5 ]
机构
[1] Nankai Univ, TEDA Sch Biol Sci & Biotechnol, TEDA, Tianjin 300457, Peoples R China
[2] Nankai Univ, Key Lab Mol Microbiol & Technol, Minist Educ, Coll Life Sci, Tianjin 300071, Peoples R China
[3] Nankai Univ, Tianjin Res Ctr Funct Genom & Biochips, TEDA Coll, Tianjin 300457, Peoples R China
[4] Nankai Univ, Tianjin Key Lab Microbial Funct Genom, TEDA Coll, TEDA, Tianjin 300457, Peoples R China
[5] Tianjin Biochip Corp, TEDA, Tianjin 300457, Peoples R China
关键词
INTERGENIC SPACER REGION; REAL-TIME PCR; ESCHERICHIA-COLI; LISTERIA-MONOCYTOGENES; SALMONELLA SPP; MULTIPLEX PCR; 16S-23S; GENE; STRAINS; DIFFERENTIATION;
D O I
10.1128/AEM.05914-11
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We established a microarray for the simultaneous detection and identification of diverse putative pathogens often associated with fishery products by targeting specific genes of Listeria monocytogenes, Salmonella, Shigella, Staphylococcus aureus, Streptococcus pyogenes, Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, and Yersinia enterocolitica and the 16S-23S rRNA gene internal transcribed spacer (ITS) region of Proteus mirabilis and Proteus vulgaris. The microarray contained 26 specific probes and was tested against a total of 123 target bacterial strains that included 55 representative strains, 68 clinical isolates, and 45 strains of other bacterial species that belonged to 8 genera and 34 species, and it was shown to be specific and reproducible. A detection sensitivity of 10 ng DNA or 10 CFU/ml for pure cultures of each target organism demonstrated that the assay was highly sensitive and reproducible. Mock and real fishery product samples were tested by the microarray, and the accuracy was 100%. The DNA microarray method described in this communication is specific, sensitive, and reliable and has several advantages over traditional methods of bacterial culture and antiserum agglutination assays.
引用
收藏
页码:8219 / 8225
页数:7
相关论文
共 32 条
[1]   Sequence diversity in the 16S-23S intergenic spacer region (ISR) of the rRNA operons in representatives of the Escherichia coli ECOR collection [J].
Antón, AI ;
Martínez-Murcia, AJ ;
Rodríguez-Valera, F .
JOURNAL OF MOLECULAR EVOLUTION, 1998, 47 (01) :62-72
[2]  
BROWN AC, 2008, UNDERSTANDING FOOD P, P53
[3]   16S-23S rDNA internal transcribed spacer regions in four Proteus species [J].
Cao, Boyang ;
Wang, Min ;
Liu, Lei ;
Zhou, Zhemin ;
Wen, Shaoping ;
Rozalski, Antoni ;
Wang, Lei .
JOURNAL OF MICROBIOLOGICAL METHODS, 2009, 77 (01) :109-118
[4]   Methodology for detection and typing of foodborne microorganisms [J].
de Boer, E ;
Beumer, RR .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 1999, 50 (1-2) :119-130
[5]  
DMITRIYEVA NF, 2002, ZH MIKROBIOL, V5, P3
[6]   A multiplex RTi-PCR reaction for simultaneous detection of Escherichia coli O157:H7, Salmonella spp. and Staphylococcus aureus on fresh, minimally processed vegetables [J].
Elizaquivel, Patricia ;
Aznar, Rosa .
FOOD MICROBIOLOGY, 2008, 25 (05) :705-713
[7]   The role of recombination and mutation in 16S-23S rDNA spacer rearrangements [J].
Gürtler, V .
GENE, 1999, 238 (01) :241-252
[8]   DNA microarray-based identification of serogroups and virulence gene patterns of Escherichia coli isolates associated with porcine postweaning diarrhea and edema disease [J].
Han, Weiqing ;
Liu, Bin ;
Cao, Boyang ;
Beutin, Lothar ;
Krueger, Ulrike ;
Liu, Hongbo ;
Li, Yayue ;
Liu, Yanqun ;
Feng, Lu ;
Wang, Lei .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (12) :4082-4088
[9]   Possible use of ail and foxA polymorphisms for detecting pathogenic Yersinia enterocolitica [J].
Huang, Ying ;
Wang, Xin ;
Cui, Zhigang ;
Yang, Yuhuan ;
Xiao, Yuchun ;
Tang, Liuying ;
Kan, Biao ;
Xu, Jianguo ;
Jing, Huaiqi .
BMC MICROBIOLOGY, 2010, 10
[10]   Listeria monocytogenes in spontaneous abortions in humans and its detection by multiplex PCR [J].
Kaur, S. ;
Malik, S. V. S. ;
Vaidya, V. M. ;
Barbuddhe, S. B. .
JOURNAL OF APPLIED MICROBIOLOGY, 2007, 103 (05) :1889-1896