摘要
      由于环境DNA分子（eDNA）在水生环境中的丰度较低，准确测定环境DNA分子中的抗生素抗性基因（ARGs）具有挑战性。在这里，我们报道了一种使用LnPO4纳米材料在水生环境中提取微量eDNA的简单且成本效益高的方法。在纳米材料中，结晶度较低的TbPO4纳米针被确定为长链DNA和短链DNA的最突出候选者，其吸附效率超过97%。用优化的洗脱液（85%PBS，15%EtOH，4g/L甘氨酸，pH 10.0）从TbPO4纳米针上洗脱吸附的DNA，最佳DNA回收率为78.83%。对于不同的环境样品，我们的方法显示出与商业提取方法相当或更好的eDNA提取效率，但成本低89%。提取的eDNA的高纯度通过高的A260/280比率来证明。利用qPCR实验，在河流样本中检测到六种常见的ARG在eDNA中的出现，其丰度范围为4.06×103至3.51×109拷贝/L。这种特定的DNA捕获器有助于评估ARGs污染的空间和时间动态，从而深入了解人类健康的潜在风险。
Abstract
Accurate determination of antibiotic resistance genes (ARGs) in environmental DNA molecules (eDNA) is challenging owing to its low abundance in the aquatic environment. Here we report a facile and cost-efficient approach to extract trace amount of eDNAs in the aquatic environment using LnPO4 nanomaterials. Among the nanomaterials, less crystalline TbPO4 nanoneedles was identified as the most prominent candidate for long stranded DNA and short stranded DNA with adsorption efficiency above 97%. The adsorbed DNA was washed off from TbPO4 nanoneedles by optimized eluant (85% PBS, 15% EtOH, 4 g/L glycine, pH 10.0) with an optimal DNA recovery of 78.83%. Our approach showed a comparable or better eDNA extraction efficiency than a commercial extraction method for different environmental samples, but 89% less cost. The high purity of the extracted eDNA was demonstrated by a high A260/280 ratio. Using qPCR experiment, the occurrence of six common ARGs in the eDNA were detected with abundance ranging from 4.06 × 103 to 3.51 × 109 copies/L in river samples. This specific DNA capturer is valuable for the evaluation of spatial and temporal dynamic of ARGs pollution to provide insight into the potential risk with regard to the human health.

https://www.sciencedirect.com/science/article/abs/pii/S0304389421021075