Genome Mining of Novel Targets and Construction of Ladder-shaped melting temperature isothermal amplification Assays for the Identification of Cronobacter sakazakii and Cronobacter malonaticus.

IF 1.9 2区农林科学 Q3 FOOD SCIENCE & TECHNOLOGY

Foodborne pathogens and disease Pub Date : 2024-09-01 Epub Date: 2024-07-04 DOI:10.1089/fpd.2024.0001

Da Ma, Ling Deng, Liangpi Huang, Aoxiang Peng, Wanglai Bi, Rui Li

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引用次数: 0

Abstract

Cronobacter species are potential pathogens that can contaminate powdered infant formula. C. sakazakii and C. malonaticus are the most common species of Cronobacter associated with infections. This study mined new molecular targets for the detection of C. sakazakii and C. malonaticus by using comparative genome approaches. Specific target genes mngB and ompR were obtained and used to detect C. sakazakii and C. malonaticus, respectively. A novel detection method, termed ladder-shape melting temperature isothermal amplification (LMTIA), was developed and evaluated. The detection limit for pure C. sakazakii DNA was 1 pg per reaction and 1 fg per reaction for C. malonaticus. The C. sakazakii, C. malonaticus, and the reference stains were all correctly identified. The amplicons can be successfully visualized and identified by naked eyes when hydroxy naphthol blue dye (HNB dye) was used in the reaction. Therefore, the LMTIA assays developed in this study showed potential application for microorganism identification and detection.

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挖掘基因组中的新靶标并构建梯形熔融温度等温扩增检测方法，以鉴定阪崎肠杆菌和丙二酸嗜盐肠杆菌。

克罗诺杆菌是可能污染婴儿配方粉的潜在病原体。阪崎杆菌和丙二酸杆菌是与感染有关的最常见的克罗诺杆菌。本研究利用比较基因组方法挖掘了检测阪崎肠杆菌和丙二酸杆菌的新分子靶标。研究获得了特异性靶基因 mngB 和 ompR，并分别用于检测阪崎肠杆菌和丙二酸杆菌。开发并评估了一种新的检测方法，称为梯形熔融温度等温扩增（LMTIA）。纯 C. sakazakii DNA 的检测限为每反应 1 pg，C. malonaticus 为每反应 1 fg。阪崎肠杆菌、丙二酸杆菌和参照染色单体都能被正确识别。在反应中使用羟基萘酚蓝染料（HNB 染料）时，可以成功地用肉眼观察和识别扩增子。因此，本研究开发的 LMTIA 检测方法在微生物鉴定和检测方面具有潜在的应用价值。

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来源期刊

Foodborne pathogens and disease 医学-食品科技

CiteScore

5.30

自引率

3.60%

发文量

审稿时长

1 months

期刊介绍： Foodborne Pathogens and Disease is one of the most inclusive scientific publications on the many disciplines that contribute to food safety. Spanning an array of issues from "farm-to-fork," the Journal bridges the gap between science and policy to reduce the burden of foodborne illness worldwide. Foodborne Pathogens and Disease coverage includes: Agroterrorism Safety of organically grown and genetically modified foods Emerging pathogens Emergence of drug resistance Methods and technology for rapid and accurate detection Strategies to destroy or control foodborne pathogens Novel strategies for the prevention and control of plant and animal diseases that impact food safety Biosecurity issues and the implications of new regulatory guidelines Impact of changing lifestyles and consumer demands on food safety.