A rapid and sensitive triplex-recombinase polymerase amplification for simultaneous differentiation of Brucella abortus, Brucella melitensi s, and Brucella suis in sera and foods.

IF 2.2 4区生物学 Q3 MICROBIOLOGY

Fems Microbiology Letters Pub Date : 2023-01-17 DOI:10.1093/femsle/fnad056

Jiang Chang, Xusen Hou, Xin Yang, Shi-Jun Zhang, De-Ying Zou, Feng Li, Ying Zhang, Yan-Song Li, Shi-Ying Lu, Pan Hu, Zeng-Shan Liu, Hong-Lin Ren

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引用次数: 0

Abstract

Brucella is the causative agent of brucellosis and can be transmitted to humans through aerosolized particles or contaminated food. Brucella abortus (B. abortus), Brucella melitensis (B. melitensis), and Brucella suis (B. suis) are the most virulent of the brucellae, but the traditional detection methods to distinguish them are time-consuming and require high instrumentation. To obtain epidemiological information on Brucella during livestock slaughter and food contamination, we developed a rapid and sensitive triplex recombinant polymerase amplification (triplex-RPA) assay that can simultaneously detect and differentiate between B. abortus, B. melitensis, and B. suis. Three pairs of primers (B1O7F/B1O7R, B192F/B192R, and B285F/B285R) were designed and screened for the establishment of the triplex-RPA assay. After optimization, the assay can be completed within 20 min at 39°C with good specificity and no cross-reactivity with five common pathogens. The triplex-RPA assay has a DNA sensitivity of 1-10 pg and a minimum detection limit of 2.14 × 104-2.14 × 105 CFU g-1 in B. suis spiked samples. It is a potential tool for the detection of Brucella and can effectively differentiate between B. abortus, B. melitensis, and B. suis S2, making it a useful tool for epidemiological investigations.

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一种快速、灵敏的三重重组酶聚合酶扩增技术，用于同时分化血清和食品中的流产布鲁氏菌、梅氏布鲁氏菌和猪布鲁氏菌。

布鲁氏菌是布鲁氏菌病的病原体，可通过雾化颗粒或受污染的食物传播给人类。流产布鲁氏菌(B. abortus)、melitensis布鲁氏菌(B. melitensis)和猪布鲁氏菌(B. suis)是布鲁氏菌中毒性最强的，但传统的检测方法既耗时又需要高仪器。为了获取畜禽屠宰和食品污染过程中布鲁氏菌的流行病学信息，我们建立了一种快速、灵敏的三重重组聚合酶扩增(triplex- rpa)方法，可以同时检测和区分abortus、B. melitensis和B. suis。设计并筛选了3对引物(b107f / b107r、B192F/B192R和B285F/B285R)，用于建立三联体rpa实验。优化后，在39℃条件下可在20 min内完成检测，特异性好，与5种常见病原菌无交叉反应。该方法的DNA灵敏度为1 ~ 10pg，最低检出限为2.14 × 104 ~ 2.14 × 105 CFU g-1。它是一种潜在的布鲁氏菌检测工具，能有效区分abortus、B. melitensis和B. suis S2，是流行病学调查的有用工具。

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来源期刊

Fems Microbiology Letters 生物-微生物学

CiteScore

4.30

自引率

0.00%

发文量

112

审稿时长

1.9 months

期刊介绍： FEMS Microbiology Letters gives priority to concise papers that merit rapid publication by virtue of their originality, general interest and contribution to new developments in microbiology. All aspects of microbiology, including virology, are covered. 2019 Impact Factor: 1.987, Journal Citation Reports (Source Clarivate, 2020) Ranking: 98/135 (Microbiology) The journal is divided into eight Sections: Physiology and Biochemistry (including genetics, molecular biology and ‘omic’ studies) Food Microbiology (from food production and biotechnology to spoilage and food borne pathogens) Biotechnology and Synthetic Biology Pathogens and Pathogenicity (including medical, veterinary, plant and insect pathogens – particularly those relating to food security – with the exception of viruses) Environmental Microbiology (including ecophysiology, ecogenomics and meta-omic studies) Virology (viruses infecting any organism, including Bacteria and Archaea) Taxonomy and Systematics (for publication of novel taxa, taxonomic reclassifications and reviews of a taxonomic nature) Professional Development (including education, training, CPD, research assessment frameworks, research and publication metrics, best-practice, careers and history of microbiology) If you are unsure which Section is most appropriate for your manuscript, for example in the case of transdisciplinary studies, we recommend that you contact the Editor-In-Chief by email prior to submission. Our scope includes any type of microorganism - all members of the Bacteria and the Archaea and microbial members of the Eukarya (yeasts, filamentous fungi, microbial algae, protozoa, oomycetes, myxomycetes, etc.) as well as all viruses.