Direct detection of thermotolerant campylobacters in chicken products by PCR and in situ hybridization
Direct detection of thermotolerant campylobacters in chicken products by PCR and in situ hybridization
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DOI：
10.1016/s0923-2508(01)01232-3
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发表时间：
2001-07-01
影响因子：
2.6
通讯作者：
Hernández, J
中科院分区：
文献类型：
--
作者：
Moreno, Y;Hernández, M;Hernández, J
我们评估了PCR和荧光原位杂交（FISH）技术在天然污染鸡肉产品中检测耐热弯曲杆菌的应用。利用16S rRNA序列数据设计两种特异性引物和寡核苷酸探针，分别用于PCR和FISH分析。PCR扩增出一个439 bp的片段，对应于耐热弯曲杆菌的特定16S RNA基因的一部分。PCR检测范围在10个细胞（富集后）到10(2)个细胞/ mL（未富集）之间变化。FISH探针能够在“加钉”和“未加钉”的自然污染样品中鉴定出耐热弯曲杆菌种类。对自然污染的样品进行PCR和FISH，并与选择培养基上分离的细胞进行比较。原位杂交技术的灵敏度低于PCR，但富集22 h后检测灵敏度显著提高。这些结果证实了基于16S rrna的技术对食品样品中弯曲杆菌的直接检测的有效性。(C) 2001年版《科学与医学》Elsevier SAS。
We have evaluated the use of PCR and fluorescent in situ hybridization (FISH) techniques for the detection of thermotolerant campylobacters in naturally contaminated chicken products. 16S rRNA sequence data was used to design two specific primers and an oligonucleotide probe for PCR and FISH analyses, respectively. The PCR protocol amplified a 439-bp fragment corresponding to a portion of specific 16S RNA gene from thermotolerant campylobacters. The detection range of the PCR assay varied between 10 cells (after enrichment) to 10(2) cells per mL (without enrichment). FISH probes were able to identify thermotolerant Campylobacter species in 'spiked' and 'unspiked' naturally contaminated samples. PCR and FISH were performed on naturally contaminated samples and compared with the isolation of cells on selective media. The in situ hybridization technique was less sensitive than PCR, although its sensitivity of detection was increased considerably after 22 h of enrichment. These results confirm the usefulness of 16S rRNA-based techniques for the direct detection of campylobacters in food samples. (C) 2001 Editions scientifiques et medicales Elsevier SAS.