Development of a multiplex PCR assay for the identification of pathogenic genes of Escherichia coli in milk and milk products

被引：22

作者：

Bottero, MT

Dalmasso, A

Soglia, D

Rosati, S

Decastelli, L

Civera, T

机构：

[1] Univ Turin, Dipartimento Patol Anim, I-10095 Grugliasco Torino, Italy

[2] Univ Turin, Dipartimento Prod Anim Epidemiol & Ecol, Turin, Italy

[3] Ist Zooprofilatt Sperimental Piemonte, Turin, Italy

来源：

MOLECULAR AND CELLULAR PROBES | 2004年 / 18卷 / 04期

关键词：

pathogenic Escherichia coli; multiplex PCR; milk; milk products;

D O I：

10.1016/j.mcp.2004.03.001

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A multiplex PCR for the simultaneous detection of some pathogenic genes of enteropathogenic, enterotoxigenic and verocytotoxin-producing Escherichia coli was developed. In this study primers found in literature as well as primers to the purpose designed were used. In this way, it was possible to generate specific fragments of 96, 170, 229, 285, 348, 414 and 510 bp for Hlya, St, EaeA, Lt, Vt1, UidA and Vt2 genes, respectively. When applied to bacterial strains experimentally inoculated in milk and milk products, the proposed PCR showed a detection limit of 5 x 10(4) CFU/ml for Hyla, St, Eaea, Vt1 primers, while for Lt and Vt2 primers the limit resulted of 106 CFU/ml. (C) 2004 Elsevier Ltd. All rights reserved.

引用

收藏

页码：283 / 288

页数：6

相关论文

共 15 条

[1] Detection of enterohemorrhagic Escherichia coli in meat foods using DNA probes, enzyme-linked immunosorbent assay and polymerase chain reaction [J].

Alexandre, M ;

Prado, V ;

Ulloa, MT ;

Arellano, C ;

Rios, M .

JOURNAL OF VETERINARY MEDICINE SERIES B-INFECTIOUS DISEASES AND VETERINARY PUBLIC HEALTH, 2001, 48 (05) :321-330

[2] BASIC LOCAL ALIGNMENT SEARCH TOOL [J].

ALTSCHUL, SF ;

GISH, W ;

MILLER, W ;

MYERS, EW ;

LIPMAN, DJ .

JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) :403-410

[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] Detection and characterization of verotoxigenic Escherichia coli by a VTEC/EHEC multiplex PCR in porcine faeces and pig carcass swabs [J].

Botteldoorn, N ;

Heyndrickx, M ;

Rijpens, N ;

Herman, L .

RESEARCH IN MICROBIOLOGY, 2003, 154 (02) :97-104

[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] ESCHERICHIA-COLI AND ENTERITIS IN ANGOLA [J].

CALUNDUNGO, R ;

MADDAU, G ;

RAPPELLI, P ;

CAPPUCCINELLI, P ;

LEITE, F ;

FRANCISCO, M ;

COLOMBO, M .

LANCET, 1994, 344 (8921) :538-539

[7] Non-O157 Shiga toxin-producing Escherichia coli infections in Europe [J].

Caprioli, A ;

Tozzi, AE ;

Rizzoni, G ;

Karch, H .

EMERGING INFECTIOUS DISEASES, 1997, 3 (04) :578-579

[8] A comparison of immunomagnetic separation and culture, RevealTM and VIPTM for the detection of E-coli O157 in enrichment cultures of naturally-contaminated raw beef, lamb and mixed meat products [J].

Chapman, PA ;

Ellin, M ;

Ashton, R .

LETTERS IN APPLIED MICROBIOLOGY, 2001, 32 (03) :171-175

[9]

Fagan PK, 1999, APPL ENVIRON MICROB, V65, P868

[10] PCR-based detection of verotoxin-producing Escherichia coli (VTEC) in ground beef [J].

Gilgen, M ;

Hübner, P ;

Höfelein, C ;

Lüthy, J ;

Candrian, U .

RESEARCH IN MICROBIOLOGY, 1998, 149 (02) :145-154