Multiplex real-time PCR method to identify Shiga toxin genes stx1 and stx2 and Escherichia coli O157:H7/H- serotype

被引：82

作者：

Jinneman, KC ^{[1
]}

Yoshitomi, KJ ^{[1
]}

Weagant, SD ^{[1
]}

机构：

[1] US FDA, Seafood Prod Res Ctr, Pacific Reg Lab NW, Bothell, WA 98021 USA

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2003年 / 69卷 / 10期

关键词：

D O I：

10.1128/AEM.69.10.6327-6333.2003

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：

A multiplex real-time PCR method to simultaneously detect the stx1 and stx2 genes of Shiga toxin-producing Escherichia coli and a unique conserved single-nucleotide polymorphism in the E. coli O157:H7/H- uidA gene has been developed. There is more than 98.6% sensitivity and 100% specificity for all three gene targets based on a panel of 138 isolates. The PCR efficiencies were greater than or equal to1.89, and as few as 6 CFU/reaction could be detected.

引用

页码：6327 / 6333

页数：7

共 36 条

[1]

Ahmed A., 1998, ESCHERICHIA COLI O15, P59

[2] Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].

Altschul, SF ;

Madden, TL ;

Schaffer, AA ;

Zhang, JH ;

Zhang, Z ;

Miller, W ;

Lipman, DJ .

NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402

[3] Rapid detection of Shiga toxin-producing bacteria in feces by multiplex PCR with molecular beacons on the smart cycler [J].

Bélanger, SD ;

Boissinot, M ;

Ménard, C ;

Picard, FJ ;

Bergeron, MG .

JOURNAL OF CLINICAL MICROBIOLOGY, 2002, 40 (04) :1436-1440

[4] Rapid detection of enterohemorrhagic Escherichia coli by real-time PCR with fluorescent hybridization probes [J].

Bellin, T ;

Pulz, M ;

Matussek, A ;

Hempen, HG ;

Gunzer, F .

JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (01) :370-374

[5] Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays [J].

Bustin, SA .

JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2000, 25 (02) :169-193

[6]

*CDCP, 1996, MMWR-MORBID MORTAL W, V45, P44

[7] SIMULTANEOUS IDENTIFICATION OF STRAINS OF ESCHERICHIA-COLI SEROTYPE O157-H7 AND THEIR SHIGA-LIKE TOXIN TYPE BY MISMATCH AMPLIFICATION MUTATION ASSAY-MULTIPLEX PCR [J].

CEBULA, TA ;

PAYNE, WL ;

FENG, P .

JOURNAL OF CLINICAL MICROBIOLOGY, 1995, 33 (01) :248-250

[8]

Centers for Disease Control and Prevention, 1997, MMWR-MORBID MORTAL W, V46, P741

[9]

Centers for Disease Control and Prevention (CDC), 1993, MMWR Morb Mortal Wkly Rep, V42, P258

[10] IDENTIFICATION OF ESCHERICHIA-COLI SEROTYPE O157-H7 BY DNA PROBE SPECIFIC FOR AN ALLELE OF UID A-GENE [J].

FENG, P .

MOLECULAR AND CELLULAR PROBES, 1993, 7 (02) :151-154

← 1 2 3 4 →