Tiet Pamela, Clark Karen C, McNamara James O, Berlin Jacob M
Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, University of Iowa , 285 Newton Road, Iowa City, Iowa 52242, United States.
Bioconjug Chem. 2017 Jan 18;28(1):183-193. doi: 10.1021/acs.bioconjchem.6b00571. Epub 2016 Dec 2.
Current water quality monitoring methods rely on growth-based measurements to detect fecal indicator bacteria, such as Escherichia coli and enterococci, and Staphylococcus aureus (S. aureus). These growth-based measurements, however, can take days to complete. This is a significant limitation in the evaluation of contaminated food and water sources. Various methods for selective in vitro detection of S. aureus have also been reported; however, these strategies, such as ELISA, agar-diffusion, PCR, or liquid chromatography-tandem mass spectrometry, all require overnight culturing or sophisticated instrumentation. There is a pressing need for a portable, simple diagnostic for S. aureus. Here, we demonstrate that oligonucleotide-functionalized gold nanoparticles (Oligo-AuNPs) can be designed to rapidly and selectively detect S. aureus with a colorimetric readout. We have functionalized a chemically modified 11-mer sequence onto AuNPs and have found that aggregation occurs in the presence of S. aureus supernantants. The particles can be stored as a lyophilized powder and reconstituted at time of use, and this has been tested in biologically relevant samples such as creek and ocean water. This approach requires minimal sample preparation and requires no extraneous instrumentation, leading to a rapid and simple diagnostic read-out that could be used in field tests to monitor food and water sources.
当前的水质监测方法依靠基于生长的测量来检测粪便指示菌,如大肠杆菌、肠球菌以及金黄色葡萄球菌(S. aureus)。然而,这些基于生长的测量可能需要数天才能完成。这在评估受污染的食物和水源时是一个重大限制。也有各种用于体外选择性检测金黄色葡萄球菌的方法被报道;然而,这些策略,如酶联免疫吸附测定(ELISA)、琼脂扩散法、聚合酶链式反应(PCR)或液相色谱 - 串联质谱法,都需要过夜培养或精密仪器。迫切需要一种用于金黄色葡萄球菌的便携式、简单诊断方法。在此,我们证明寡核苷酸功能化金纳米颗粒(Oligo - AuNPs)可被设计用于通过比色读数快速且选择性地检测金黄色葡萄球菌。我们已将化学修饰的11聚体序列功能化到金纳米颗粒上,并且发现金黄色葡萄球菌上清液存在时会发生聚集。这些颗粒可以作为冻干粉末储存并在使用时复溶,并且已在诸如溪水和海水等生物相关样本中进行了测试。这种方法所需的样品制备最少,且不需要额外的仪器,从而实现快速且简单的诊断读数,可用于现场测试以监测食物和水源。