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利用诱饵捕获技术进行靶向宏基因组学分析以检测零售肉类和海鲜中的抗生素抗性基因。

Targeted metagenomics using bait-capture to detect antibiotic resistance genes in retail meat and seafood.

作者信息

Flint Annika, Cooper Ashley, Rao Mary, Weedmark Kelly, Carrillo Catherine, Tamber Sandeep

机构信息

Bureau of Microbial Hazards, Health Canada, Sir Frederick Banting Driveway, Ottawa, ON, Canada.

Research and Development, Ottawa Laboratory (Carling), Canadian Food Inspection Agency, Ottawa, ON, Canada.

出版信息

Front Microbiol. 2023 Jul 13;14:1188872. doi: 10.3389/fmicb.2023.1188872. eCollection 2023.

Abstract

Metagenomics analysis of foods has the potential to provide comprehensive data on the presence and prevalence of antimicrobial resistance (AMR) genes in the microbiome of foods. However, AMR genes are generally present in low abundance compared to other bacterial genes in the food microbiome and consequently require multiple rounds of in-depth sequencing for detection. Here, a metagenomics approach, using bait-capture probes targeting antimicrobial resistance and plasmid genes, is used to characterize the resistome and plasmidome of retail beef, chicken, oyster, shrimp, and veal enrichment cultures ( = 15). Compared to total shotgun metagenomics, bait-capture required approximately 40-fold fewer sequence reads to detect twice the number of AMR gene classes, AMR gene families, and plasmid genes across all sample types. For the detection of critically important extended spectrum beta-lactamase (ESBL) genes the bait capture method had a higher overall positivity rate (44%) compared to shotgun metagenomics (26%), and a culture-based method (29%). Overall, the results support the use of bait-capture for the identification of low abundance genes such as AMR genes from food samples.

摘要

食品宏基因组学分析有潜力提供关于食品微生物组中抗菌药物耐药性(AMR)基因的存在及流行情况的全面数据。然而,与食品微生物组中的其他细菌基因相比,AMR基因通常以低丰度存在,因此需要多轮深度测序才能检测到。在此,采用一种宏基因组学方法,使用靶向抗菌药物耐药性和质粒基因的捕获探针,来表征零售牛肉、鸡肉、牡蛎、虾和小牛肉富集培养物(n = 15)的耐药基因组和质粒组。与全基因组鸟枪法宏基因组学相比,捕获探针法检测所有样本类型中的AMR基因类别、AMR基因家族和质粒基因数量翻倍时所需的序列读数大约少40倍。对于检测至关重要的超广谱β-内酰胺酶(ESBL)基因,捕获探针法的总体阳性率(44%)高于全基因组鸟枪法宏基因组学(丝26%)和基于培养的方法(29%)。总体而言,研究结果支持使用捕获探针法来鉴定食品样本中低丰度基因,如AMR基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5b0/10373929/b2f32f266762/fmicb-14-1188872-g001.jpg

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