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通过高分辨率熔解曲线分析鉴定和区分弯曲杆菌属菌种。

Identification and differentiation of Campylobacter species by high-resolution melting curve analysis.

作者信息

Hoseinpour Fatemeh, Foroughi Azadeh, Nomanpour Bizhan, Nasab Rezvan Sobhani

机构信息

Department of Pathobiology & Basic Sciences, Faculty of Veterinary Medicine, Razi University, Kermanshah, Iran.

Department of Microbiology, Kermanshah University of Medical Sciences, Kermanshah, Iran.

出版信息

Microb Pathog. 2017 Jul;108:109-113. doi: 10.1016/j.micpath.2017.05.009. Epub 2017 May 3.

Abstract

BACKGROUND

Campylobacter jejuni and Campylobacter coli are the important food-born zoonotic pathogen, also are leading causes of human food borne illnesses worldwide. cadF gene is expressed in all C. jejuni and C. coli strains and mediates cell binding to the cell matrix protein, Fibronectin. High-resolution melting (HRM) analysis is emerging as an efficient and robust method for discriminating DNA sequence variants. The goal of this study was to apply HRM analysis to identification of C. jejuni and C. coli.

MATERIALS AND METHODS

A total of 100 samples were obtained from chicken in Kermanshah, Iran. HRM analysis based on cadF gene and Eva Green was developed to the identification of Campylobacter to the species level.

RESULTS

Fifty-five of 100 samples were positive as campylobacter (7 C. jejuni, 29 C. coli, 16 mixes and 3 unknown). Minor variations were observed in melting point temperatures of C. coli or C. jejuni isolates and this variation can be used to the differentiation between C. coli or C. jejuni isolates.

CONCLUSIONS

The results of this study indicated that HRM curve analysis can be a suitable technique and rapid technique for distinguishing between C. jejuni and C. coli isolates.

摘要

背景

空肠弯曲菌和结肠弯曲菌是重要的食源性人畜共患病原体,也是全球人类食源性疾病的主要病因。cadF基因在所有空肠弯曲菌和结肠弯曲菌菌株中均有表达,并介导细胞与细胞基质蛋白纤连蛋白的结合。高分辨率熔解(HRM)分析正成为一种鉴别DNA序列变异的高效且可靠的方法。本研究的目的是应用HRM分析来鉴定空肠弯曲菌和结肠弯曲菌。

材料与方法

从伊朗克尔曼沙赫的鸡肉中总共获取了100份样本。基于cadF基因和Eva Green开发了HRM分析方法,用于将弯曲菌鉴定到种水平。

结果

100份样本中有55份弯曲菌检测呈阳性(7份空肠弯曲菌、29份结肠弯曲菌、16份混合菌和3份未知菌)。在结肠弯曲菌或空肠弯曲菌分离株的熔点温度上观察到微小差异,这种差异可用于区分结肠弯曲菌或空肠弯曲菌分离株。

结论

本研究结果表明,HRM曲线分析可以是一种适用于区分空肠弯曲菌和结肠弯曲菌分离株的快速技术。

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