这项研究评估了十种核酸提取方案(EP1至EP10),用于测量四个飞机废水样品(AWW1至AWW4)中的五个内源性抗生素抗性基因(ARG)。目标ARGs,包括BlaCTX-M,blaNDM-1,ermB,qnrS,还有tetA,包含高度和最低限度的ARGs。使用DNeasy血液和组织试剂盒和AllPrepPowerViralDNA/RNA试剂盒,在四个飞机废水样品中始终检测到TetA和ermB。QnrS在特定的提取方案和等分体积下显示出高检测率。ARGs的浓度因飞机废水样本而异,不同的提取方案影响定量结果。tetA的浓度,ermB,和AWW1中的qnrS是不同的,而AWW2到AWW4表现出更广泛的tetA范围,ermB,qnrS,BlaCTX-M,和blaNDM-1.EP1始终产生几种ARG的最高浓度。集体数据分析显示,在十种提取方案中,ARG浓度各不相同,这表明在飞机废水样品的ARG监测中仔细选择提取方案的重要性。根据结果,我们建议,小样品体积(低至0.2mL)可能足以用于飞机废水样品中的ARG表征。该发现还强调需要考虑在不损害核酸提取效率的情况下去除卫生纸。该研究强调了ARGs飞机废水监测的前景,呼吁进一步调查独特ARG通过交通枢纽的进口和传播。
This study evaluated ten nucleic acid extraction protocols (EP1 to EP10) for measuring five endogenous antibiotic resistance genes (ARGs) in four
aircraft wastewater samples (AWW1 to AWW4). The targeted ARGs, including blaCTX-M, blaNDM-1, ermB, qnrS, and tetA, encompassed highly and minimally abundant ARGs. TetA and ermB were consistently detected across four
aircraft wastewater samples using the DNeasy Blood and Tissue Kit and the AllPrep PowerViral DNA/RNA kit. QnrS displayed high detection rates with specific extraction protocols and aliquot volumes. Concentrations of ARGs varied across
aircraft wastewater samples, with differing extraction protocols influencing quantitative results. The concentrations of tetA, ermB, and qnrS in AWW1 were distinct, while AWW2 to AWW4 exhibited a broader range for tetA, ermB, qnrS, blaCTX-M, and blaNDM-1. EP1 consistently produced the highest concentrations for several ARGs. Collective data analysis revealed varying ARG concentrations across the ten extraction protocols, suggesting the importance of careful extraction protocol selection in ARG monitoring in
aircraft wastewater samples. Based on the results, we suggest that a small sample volume (as low as 0.2 mL) may be sufficient for ARG characterization in aircraft wastewater samples. The findings also emphasize the need for considering toilet paper removal without compromising nucleic acid extraction efficiency. The study highlights promising prospects for
aircraft wastewater monitoring of ARGs, calling for further investigation into the import and spread of unique ARGs through transport hubs.