A survey on the incidence of Campylobacter spp. and the development of a surface adhesion polymerase chain reaction (SA-PCR) assay for the detection of Campylobacter jejuni in retail meat products

被引:31
|
作者
Cloak, OM
Duffy, G
Sheridan, JJ
Blair, IS
McDowell, DA
机构
[1] TEAGASC, Natl Food Ctr, Dublin 15, Ireland
[2] Univ Ulster, Jordanstown BT37 0QB, Antrim, North Ireland
关键词
D O I
10.1006/fmic.2001.0400
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 rapid and sensitive method for the detection of Campylobacter in meat products was developed. Campylobacter were isolated from a range of Irish retail meats (n = 80) and poultry (n = 100) samples by direct plating on Campylobacter Selective Agar (CCDA, Oxoid). A total of 30.5% of samples tested positive for Campylobacter and Campylobacter jejuni was the most commonly identified species. The pathogen was defected in 39 (65%) poultry, 12 (60%) offal and 4 (20%) minced beef samples examined: Estimates derived from direct plate counts ranged from log(10) 0.7 to log(10) 2.75 cfu g(-1) The data from this study was used in the development of a rapid and sensitive method for the detection of Campylobacter in poultry. A rapid method was developed based on an initial sample enrichment for 24 h in Campylobacter Enrichment Broth (CEB) recovery of the pathogen from the enriched sample by surface adhesion onto a polycarbonate membrane, phenol:chloroform extraction of DNA from she ad herent bacteria, and PCR analysis using primers specific for the flagellin A gene (present in C, jejuni and C. coli). The developed surface adhesion PCR (SA-PCR) technique had a detection limit of log(10) 4-5 and could be completed within 29h. Results from SA-PCR analysis of a number of retail samples (n=50) correlated favourably with traditional plate culture results, i.e. 34 samples were found to contain Campylobacter by both methods. (C) 2001 Academic Press.
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收藏
页码:287 / 298
页数:12
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