用于检测和区分鸡肉和羊肉中的大肠杆菌、胎生大肠杆菌和洁净大肠杆菌与其他弯曲杆菌的四重 qPCR。

IF 2.1 4区 生物学 Q3 MICROBIOLOGY Brazilian Journal of Microbiology Pub Date : 2024-09-01 Epub Date: 2024-07-09 DOI:10.1007/s42770-024-01437-4
Marwan Abu-Halaweh, Eman Al-Bsoul
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引用次数: 0

摘要

弯曲杆菌是革兰氏阴性菌,被认为是从家禽样本中分离出来的主要菌属,与肠胃炎有关。由于传统的培养方法耗时长、技术要求高,因此需要一种快速可行的方法来鉴定和区分与之密切相关的菌属,包括大肠弯曲杆菌、胎儿弯曲杆菌和空肠弯曲杆菌。本研究采用基于培养和预富集的 Quadraplex 实时 PCR 技术,针对大肠弯曲杆菌、胎儿弯曲杆菌、空肠弯曲杆菌和弯曲杆菌属的 OrfA、CstA、HipO 和 16 S rRNA 基因,对鸡肉和羊肉样本(n = 125)进行了分析。通过培养和实时 PCR 对 125 份鸡肉和羊肉样本进行的分析表明,两种方法的结果高度一致。本研究显示弯曲杆菌的流行率很高(鸡肉和羊肉中的流行率分别为 35% 和 32%),其中空肠弯曲杆菌的流行率最高。反应效率在 90% 到 110% 之间,空肠弯曲菌的检出率低至 8.9 fg。使用所述 Quadraplex 实时 PCR 方法可满足对羊肉和鸡肉中快速检测和鉴别方法的需求。
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Quadruplex qPCR for detection and discrimination of C. Coli,C. fetus, and C. Jejuni from other Campylobacter species in chicken and sheep meat.

Campylobacter is gram-negative bacteria considered the predominant genera isolated from poultry samples and associated with gastroenteritis. Due to the problems in conventional cultural methods of time-consuming and technically demanding requirements, a rapid and feasible method for their identification and discrimination of the closely related spp. Including Campylobacter coli, Campylobacter fetus, and Campylobacter jejuni is needed. This study analyzes the chicken and sheep meats samples (n = 125) using culture and pre-enrichment-based Quadraplex real-time PCR by targeting OrfA, CstA, HipO, and 16 S rRNA genes of C. coli, C. fetus, C. jejuni and Campylobacter spp. Respectively. The analysis of 125 chicken and sheep meat samples by culture and real-time PCR showed high concordance between the results of the two methods. The present study show high prevalence of Campylobacter species (35% and 32% from chicken and meat respectively) of which C. jejuni were the most abundant. Reaction efficiencies were between 90 and 110%, and detect as low as 8.9 fg in C. jejuni. The need for quick detection and discrimination methods in sheep and chicken meat can be met using the described Quadraplex real-time PCR methodology.

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来源期刊
Brazilian Journal of Microbiology
Brazilian Journal of Microbiology 生物-微生物学
CiteScore
4.10
自引率
4.50%
发文量
216
审稿时长
1.0 months
期刊介绍: The Brazilian Journal of Microbiology is an international peer reviewed journal that covers a wide-range of research on fundamental and applied aspects of microbiology. The journal considers for publication original research articles, short communications, reviews, and letters to the editor, that may be submitted to the following sections: Biotechnology and Industrial Microbiology, Food Microbiology, Bacterial and Fungal Pathogenesis, Clinical Microbiology, Environmental Microbiology, Veterinary Microbiology, Fungal and Bacterial Physiology, Bacterial, Fungal and Virus Molecular Biology, Education in Microbiology. For more details on each section, please check out the instructions for authors. The journal is the official publication of the Brazilian Society of Microbiology and currently publishes 4 issues per year.
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