Evaluation of PCR for detection of Campylobacter in a national broiler surveillance programme in Denmark

被引:37
作者
Lund, M [1 ]
Wedderkopp, A [1 ]
Waino, M [1 ]
Nordentoft, S [1 ]
Bang, DD [1 ]
Pedersen, K [1 ]
Madsen, M [1 ]
机构
[1] Danish Vet Inst, DK-8200 Aarhus N, Denmark
关键词
broiler; Campylobacter; DNA isolation; PCR;
D O I
10.1046/j.1365-2672.2003.01934.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
Aims: To develop and evaluate a rapid and sensitive PCR method for detection of Campylobacter spp. directly from chicken faeces. Methods and Results: DNA was isolated from faecal swabs using magnetic beads followed by PCR using a prealiquoted PCR mixture, which had been stored in the freezer. The result could be obtained in <6 h. The method was evaluated on 1282 samples from the Danish surveillance programme for Campylobacter in broilers by comparing with conventional culture. The diagnostic specificity was calculated to be 0.99. The detection limits of the PCR method and of the conventional culture were compared using spiked control material. For both methods the detection limit was 36 CFU ml(-1) . Conclusions: It was concluded that the PCR proved useful for detection of Campylobacter in pooled cloacal swabs from broilers. Significance and Impact of the Study: By taking cloacal samples in the broiler flocks the technique can be used as an important tool for planning and directing the broiler slaughtering process. This will be a great help in minimizing the risk of contaminating Campylobacter-free flocks at the abattoir.
引用
收藏
页码:929 / 935
页数:7
相关论文
共 42 条
  • [1] Abu Al-Soud W, 1998, APPL ENVIRON MICROB, V64, P3748
  • [2] Development of a PCR assay suitable for Campylobacter spp. mass screening programs in broiler production
    Bang, DD
    Pedersen, K
    Madsen, M
    [J]. JOURNAL OF RAPID METHODS AND AUTOMATION IN MICROBIOLOGY, 2001, 9 (02) : 97 - 113
  • [3] Use of magnetic beads for tissue DNA extraction and IS6110 Mycobacterium tuberculosis PCR
    Caldarelli-Stefano, R
    Vago, L
    Bonetto, S
    Nebuloni, M
    Costanzi, G
    [J]. JOURNAL OF CLINICAL PATHOLOGY-MOLECULAR PATHOLOGY, 1999, 52 (03): : 158 - 160
  • [4] Detection of Campylobacter jejuni in poultry samples using an enzyme-linked immunoassay coupled with an enzyme electrode
    Che, YH
    Li, YB
    Slavik, M
    [J]. BIOSENSORS & BIOELECTRONICS, 2001, 16 (9-12) : 791 - 797
  • [5] Rapid isolation of PCR-ready DNA from blood, bone marrow and cultured cells, based on paramagnetic beads
    Deggerdal, A
    Larsen, F
    [J]. BIOTECHNIQUES, 1997, 22 (03) : 554 - 557
  • [6] Campylobacter contamination in French chicken production from farm to consumers.: Use of a PCR assay for detection and identification of Campylobacter jejuni and Camp. coli
    Denis, M
    Refrégier-Petton, J
    Laisney, MJ
    Ermel, G
    Salvat, G
    [J]. JOURNAL OF APPLIED MICROBIOLOGY, 2001, 91 (02) : 255 - 267
  • [7] The magnetic immuno-polymerase chain reaction assay for the detection of Campylobacter in milk and poultry
    Docherty, L
    Adams, MR
    Patel, P
    McFadden, J
    [J]. LETTERS IN APPLIED MICROBIOLOGY, 1996, 22 (04) : 288 - 292
  • [8] DISCRIMINATION AMONG THERMOPHILIC CAMPYLOBACTER SPECIES BY POLYMERASE CHAIN-REACTION AMPLIFICATION OF 23S-RIBOSOMAL-RNA GENE FRAGMENTS
    EYERS, M
    CHAPELLE, S
    VANCAMP, G
    GOOSSENS, H
    DEWACHTER, R
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1993, 31 (12) : 3340 - 3343
  • [9] DISCRIMINATION AMONG THERMOPHILIC CAMPYLOBACTER SPECIES BY POLYMERASE CHAIN-REACTION AMPLIFICATION OF 23S RIBOSOMAL-RNA GENE FRAGMENTS (VOL 31, PG 3341, 1994)
    EYERS, M
    CHAPELLE, S
    VANCAMP, G
    GOOSSENS, H
    DEWACHTER, R
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 1994, 32 (06) : 1623 - 1623
  • [10] Fermér C, 1999, J CLIN MICROBIOL, V37, P3370