MRG 芯片：一种基于高通量 qPCR 的重金属（样体）抗性评估工具,Environmental Science & Technology

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MRG 芯片：一种基于高通量 qPCR 的重金属（样体）抗性评估工具
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2022-07-25 , DOI: 10.1021/acs.est.2c00488
Jiaojiao Zhu _{1,

2} , Qiong Huang _{1,

2} , Xinyi Peng _{1,

2} , Xinyuan Zhou ₃ , Shenghan Gao _{1,

2} , Yuanping Li ₄ , Xuesong Luo _{1,

2} , Yi Zhao ₅ , Christopher Rensing ₄ , Jianqiang Su ₃ , Peng Cai _{1,

2,

6} , Yurong Liu _{1,

2,

6} , Wenli Chen ₁ , Xiuli Hao _{1,

2,

6} , Qiaoyun Huang _{1,

2,

6}

Affiliation

几个世纪以来，细菌金属解毒机制在纯培养系统中得到了很好的研究。然而，由于缺乏全面可靠的量化工具，在社区层面分析金属电阻决定因素仍然是一个挑战。在这里，一种新型的高通量定量聚合酶链反应 (HT-qPCR) 芯片，称为金属抗性基因 (MRG) 芯片，已被开发用于量化参与 9 种金属稳态的基因。MRG芯片包含77个新设计的简并引物组和9个已发表的引物组，涵盖56个金属抗性基因。分类覆盖的计算评估表明，MRG芯片具有广泛的覆盖范围，匹配2个界、29个门、64个纲、130个目、226个科和382个属。R ² > 0.993。HT-qPCR 的计算评估和熔解曲线分析均验证了高特异性。MRG 芯片已成功应用于表征各种金属电阻决定因素在自然和人类相关环境中的分布，证实了其广泛的应用范围。总的来说，MRG 芯片是一种功能强大且高效的高通量定量工具，用于探索微生物金属耐药性。

"点击查看英文标题和摘要"

MRG Chip: A High-Throughput qPCR-Based Tool for Assessment of the Heavy Metal(loid) Resistome

Bacterial metal detoxification mechanisms have been well studied for centuries in pure culture systems. However, profiling metal resistance determinants at the community level is still a challenge due to the lack of comprehensive and reliable quantification tools. Here, a novel high-throughput quantitative polymerase chain reaction (HT-qPCR) chip, termed the metal resistance gene (MRG) chip, has been developed for the quantification of genes involved in the homeostasis of 9 metals. The MRG chip contains 77 newly designed degenerate primer sets and 9 published primer sets covering 56 metal resistance genes. Computational evaluation of the taxonomic coverage indicated that the MRG chip had a broad coverage matching 2 kingdoms, 29 phyla, 64 classes, 130 orders, 226 families, and 382 genera. Temperature gradient PCR and HT-qPCR verified that 57 °C was the optimal annealing temperature, with amplification efficiencies of over 94% primer sets achieving 80–110%, with R² > 0.993. Both computational evaluation and the melting curve analysis of HT-qPCR validated a high specificity. The MRG chip has been successfully applied to characterize the distribution of diverse metal resistance determinants in natural and human-related environments, confirming its wide scope of application. Collectively, the MRG chip is a powerful and efficient high-throughput quantification tool for exploring the microbial metal resistome.

更新日期：2022-07-25

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