MULTIPLEX PCR FOR THE DETECTION OF CAMPYLOBACTER SPP. AND SALMONELLA SPP. IN CHICKEN MEAT

被引:21
作者
Alves, Juliane [1 ]
Marques, Viviani Vieira [2 ]
Protasio Pereira, Luiz Filipe [3 ]
Hirooka, Elisa Yoko [1 ]
Rocha Moreira De Oliveira, Tereza Cristina [1 ]
机构
[1] Univ Estadual Londrina, Dept Food Sci & Technol, BR-86051990 Londrina, Parana, Brazil
[2] IAPAR, Londrina, Parana, Brazil
[3] Embrapa Cafe, Agron Inst Parana, Biotechnol Lab, Londrina, Parana, Brazil
关键词
POLYMERASE-CHAIN-REACTION; ESCHERICHIA-COLI O157; LISTERIA-MONOCYTOGENES; ASSAY; IDENTIFICATION; AMPLIFICATION;
D O I
10.1111/j.1745-4565.2012.00386.x
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
The control of Salmonella spp. and Campylobacter spp. in chicken meat is essential for avoiding sanitary barriers and preventing human disease. The aim of this study was to develop a multiplex polymerase chain reaction assay (mPCR) for the rapid detection of these bacteria in raw chicken meat. The mPCR was developed using the Styinva-JHO-Right and Styinva-JHO-Left primers (specific for Salmonella spp.) and the OT1559 and 18-1 primers (specific for Campylobacter spp.). The specificity of the assay was 100% and it was able to detect 102 cfu/mL of Campylobacter spp. after the selective enrichment and 1 cfu/mL of Salmonella spp. after nonselective enrichment. Fifty raw chicken meat samples were analyzed; 4% were contaminated with Salmonella spp. and 56% with Campylobacter spp. The results obtained using mPCR were confirmed by conventional culturing methods. The developed mPCR method is a relatively inexpensive and efficient means to detect these bacteria after 24 h of enrichment. PRACTICAL APPLICATIONS The developed multiplex polymerase chain reaction method (mPCR) is a relatively inexpensive and efficient means to detect Salmonella spp. and Campylobacter spp. in chicken meat after 24 h of enrichment. The detection of these pathogens in a few hours allows the food supply chain to take appropriate measures quickly to prevent the distribution of contaminated food. Rapid and simultaneous detection of these bacteria in chicken meat can assist in the implementation of the preventive measures that can reduce contamination, which is very useful for Brazil, the third largest producer of chicken meat and the largest exporter of this product. Moreover, the developed mPCR may speed up the identification of suspected colonies of those bacteria on the selective media used in conventional culture methods.
引用
收藏
页码:345 / 350
页数:6
相关论文
共 27 条
[1]  
Abolmaaty A, 2000, MICROBIOS, V101, P181
[2]   POLYMERASE CHAIN-REACTION IN FOOD MICROBIOLOGY [J].
CANDRIAN, U .
JOURNAL OF MICROBIOLOGICAL METHODS, 1995, 23 (01) :89-103
[3]   Identification of Salmonella spp. isolates from chicken abattoirs by multiplex-PCR [J].
Cortez, A. L. L. ;
Carvalho, A. C. F. B. ;
Ikuno, A. A. ;
Burger, K. P. ;
Vidal-Martins, A. M. C. .
RESEARCH IN VETERINARY SCIENCE, 2006, 81 (03) :340-344
[4]   PCR multiplex for detection of Salmonella Enteritidis, Typhi and Typhimurium and occurrence in poultry meat [J].
de Freitas, Camila Guimaraes ;
Santana, Angela Patricia ;
Caldeira da Silva, Patricia Helena ;
Picao Goncalves, Vitor Salvador ;
Ferreira Barros, Marcia de Aguiar ;
Goncalves Torres, Fernando Araripe ;
Murata, Luci Sayori ;
Perecmanis, Simone .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 2010, 139 (1-2) :15-22
[5]   The magnetic immuno-polymerase chain reaction assay for the detection of Campylobacter in milk and poultry [J].
Docherty, L ;
Adams, MR ;
Patel, P ;
McFadden, J .
LETTERS IN APPLIED MICROBIOLOGY, 1996, 22 (04) :288-292
[6]   Improved Sample Preparation for Real-Time PCR Detection of Listeria monocytogenes in Hot-Smoked Salmon using Filtering and Immunomagnetic Separation Techniques [J].
Duodu, Samuel ;
Mehmeti, Ibrahim ;
Holst-Jensen, Arne ;
Loncarevic, Semir .
FOOD ANALYTICAL METHODS, 2009, 2 (01) :23-29
[7]   Twelve hour real-time PCR technique for the sensitive and specific detection of Salmonella in raw and ready-to-eat meat products [J].
Ellingson, JLE ;
Anderson, JL ;
Carlson, SA ;
Sharma, VK .
MOLECULAR AND CELLULAR PROBES, 2004, 18 (01) :51-57
[8]   Simultaneous detection of Escherichia coli O175:H7, Salmonella spp., and Listeria monocytogenes by multiplex PCR [J].
Germini, Andrea ;
Masola, Annalisa ;
Carnevali, Paola ;
Marchelli, Rosangela .
FOOD CONTROL, 2009, 20 (08) :733-738
[9]   Automated 5′ nuclease PCR assay for identification of Salmonella enterica [J].
Hoorfar, J ;
Ahrens, P ;
Rådström, P .
JOURNAL OF CLINICAL MICROBIOLOGY, 2000, 38 (09) :3429-3435
[10]   Simultaneous detection of Listeria monocytogenes and Salmonella by multiplex PCR in cooked ham [J].
Jofré, A ;
Martin, B ;
Garriga, M ;
Hugas, M ;
Pla, M ;
Rodríguez-Lázaro, D ;
Aymerich, T .
FOOD MICROBIOLOGY, 2005, 22 (01) :109-115