A New Real-Time PCR Assay for the Specific Detection of Salmonella spp. Targeting the bipA Gene

被引：39

作者：

Calvo, Laia ^{[1
]}

Martinez-Planells, Asuncion ^{[2
]}

Pardos-Bosch, Joana ^{[2
]}

Garcia-Gil, L. Jesus ^{[1
]}

机构：

[1] Univ Girona, Lab Clin Microbiol & Infect Dis, E-17071 Girona, Spain

[2] Serv Terr Salut Girona, Lab Salut Publ, E-17004 Girona, Spain

来源：

FOOD ANALYTICAL METHODS | 2008年 / 1卷 / 04期

关键词：

Salmonella spp; BipA; Real-time PCR; Food Analysis;

D O I：

10.1007/s12161-007-9008-x

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

A straightforward real-time polymerase chain reaction (PCR)-based assay was designed and evaluated for the detection of Salmonella spp. in food and water samples. This now assay is based on the specific detection of the bipA gene of Salmonella, which encodes a protein of the guanosine triphosphate (GTP)-binding elongation family that displays global modulating properties, by regulating a wide variety of downstream processes. The new method correctly identified all 48 Salmonella strains used in the inclusivity test, and did not detect all 30 non-Salmonella species tested. The method was evaluated by analyzing 120 diverse food and water samples enriched in buffered peptone water. The bipA-based real-time PCR assay showed 100% efficiency, sensitivity, and specificity compared to the invA-based method previously published, which was developed as a part of a European project for the standardization of PCR methods in food microbiology. The assay includes an independent internal amplification control (IAC) in each reaction to control false negative results.

引用

页码：236 / 242

页数：7

共 26 条

[1]

ALTSCHUL SF, 1997, NUCLEIC ACIDS RES, V25, P3402

[2] Formate protects stationary-phase Escherichia coli and Salmonella cells from killing by a cationic antimicrobial peptide [J].

Barker, HC ;

Kinsella, N ;

Jaspe, A ;

Friedrich, T ;

O'Connor, CD .

MOLECULAR MICROBIOLOGY, 2000, 35 (06) :1518-1529

[3] Rapid identification of Salmonella serovars in feces by specific detection of virulence genes, invA and spvC, by an enrichment broth culture-multiplex PCR combination assay [J].

Chiu, CH ;

Ou, JT .

JOURNAL OF CLINICAL MICROBIOLOGY, 1996, 34 (10) :2619-2622

[4] PCR amplification of the fimA gene sequence of Salmonella typhimurium, a specific method for detection of Salmonella spp [J].

Cohen, HJ ;

Mechanda, SM ;

Lin, W .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1996, 62 (12) :4303-4308

[5] BipA:: a tyrosine-phosphorylated GTPase that mediates interactions between enteropathogenic Escherichia coli (EPEC) and epithelial cells [J].

Farris, M ;

Grant, A ;

Richardson, TB ;

O'Connor, CD .

MOLECULAR MICROBIOLOGY, 1998, 28 (02) :265-279

[6] Visualization of pathogenicity regions in bacteria [J].

Friis, C ;

Jensen, LJ ;

Ussery, DW .

GENETICA, 2000, 108 (01) :47-51

[7]

GALAN JE, 1992, J BACTERIOL, V174, P4338

[8] Co-ordination of pathogenicity island expression by the BipA GTPase in enteropathogenic Escherichia coli (EPEC) [J].

Grant, AJ ;

Farris, M ;

Alefounder, P ;

Williams, PH ;

Woodward, MJ ;

O'Connor, CD .

MOLECULAR MICROBIOLOGY, 2003, 48 (02) :507-521

[9] Selective amplification of tyv (rfbE), prt (rfbS), viaB, and fliC genes by multiplex PCR for identification of Salmonella enterica [J].

Hirose, K ;

Itoh, K ;

Nakajima, H ;

Kurazono, T ;

Yamaguchi, M ;

Moriya, K ;

Ezaki, T ;

Kawamura, Y ;

Tamura, K ;

Watanabe, H .

JOURNAL OF CLINICAL MICROBIOLOGY, 2002, 40 (02) :633-636

[10] Making internal amplification control mandatory for diagnostic PCR [J].

Hoorfar, J ;

Cook, N ;

Malorny, B ;

Wagner, M ;

De Medici, D ;

Abdulmaajood, A ;

Fach, P .

JOURNAL OF CLINICAL MICROBIOLOGY, 2003, 41 (12) :5835-5835

← 1 2 3 →