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用于检测禽肉中结肠弯曲杆菌和空肠弯曲杆菌的聚合酶链反应测定法。

Polymerase chain reaction assay for detection of Campylobacter coli and Campylobacter jejuni in poultry meat.

作者信息

Manzano M, Pipan C, Botta G, Comi G

机构信息

Dipartimento di Scienze degli Alimenti, Università di Udine, Italy.

出版信息

Zentralbl Hyg Umweltmed. 1995 Jun;197(5):370-86.

PMID:8672221
Abstract

Primers of 16S rRNA and flaA genes were used to optimise a PCR technique for detecting thermotolerant campylobacters in poultry meat. Different methods for crude DNA extraction were also evaluated. The use of flaA primers and extraction of nucleic acid by boiling and proteinase K gave good results in the detection of Campylobacter either in artificially or naturally contaminated foodstuffs. The lowest sensitivity limit for the PCR reaction was 10(1)-10(2) thermophilic Campylobacter cells either in pure cultures or in artificially and naturally contaminated poultry skins, corresponding to a concentration of 10(2)-10(3) Campylobacter/ml or g product. The PCR method we devised had a high sensitivity and specificity. It appears to give better results than conventional methods and is very easy and fast, requiring only eight hours to detect thermotolerant Campylobacter from poultry meat. In contrast, conventional methods require almost 4 days.

摘要

使用16S rRNA和flaA基因引物优化了一种用于检测禽肉中耐热弯曲杆菌的PCR技术。还评估了不同的粗DNA提取方法。使用flaA引物并通过煮沸和蛋白酶K提取核酸,在检测人工或天然污染食品中的弯曲杆菌方面取得了良好效果。PCR反应的最低灵敏度极限为纯培养物中或人工和天然污染的禽皮中的10(1)-10(2)个嗜热弯曲杆菌细胞,相当于10(2)-10(3)个弯曲杆菌/毫升或克产品的浓度。我们设计的PCR方法具有高灵敏度和特异性。它似乎比传统方法效果更好,而且非常简便快捷,从禽肉中检测耐热弯曲杆菌仅需8小时。相比之下,传统方法几乎需要4天。

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