Veigas B, Fortunato E, Baptista P V
CIGMH, Departamento de Ciências da Vida, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus de Caparica, 2829-516, Caparica, Portugal.
Methods Mol Biol. 2015;1256:41-56. doi: 10.1007/978-1-4939-2172-0_3.
Nanotechnology based diagnostics has provided improved tools for pathogen detection and sensitive and specific characterization of antibiotic resistance signatures. Tuberculosis (TB) is caused by members of the Mycobacterium tuberculosis Complex (MTBC) and, according to the World Health Organization, is one of the most serious infectious diseases in the world. Recent advances in molecular diagnostics of TB have improved both the detection time and sensitivity but they still require specialized technical personnel and cumbersome laboratory equipment. Diagnostics at point-of-need is crucial to TB control as it may provide rapid identification of pathogen together with the resistance profile of TB strains, originated from single nucleotide polymorphisms (SNPs) in different loci, allowing for a more accurate indication of the adequate therapy.Gold nanoparticles have been widely used in molecular diagnostics platforms. Here, we describe the use of gold nanoprobes (oligonucleotide functionalized gold nanoparticles) to be used in a non-cross-linking colorimetric method for the direct detection of specific DNA targets. Due to the remarkable optical properties of gold nanoparticles, this detection system provides colorimetric detection of the pathogen together with the potential of identification of several single nucleotide polymorphisms (SNPs) involved in TB resistance to antibiotics. For point-of-need use, we adapted this strategy to a low-cost mobile scheme using a paper based revelation platform and where the spectral signature is transposed to RGB data via a smartphone device. This way, identification of pathogen and characterization of resistance signatures is achieved at point-of-need.
基于纳米技术的诊断方法为病原体检测以及抗生素耐药特征的灵敏和特异性表征提供了改进的工具。结核病(TB)由结核分枝杆菌复合群(MTBC)的成员引起,据世界卫生组织称,它是世界上最严重的传染病之一。结核病分子诊断的最新进展改善了检测时间和灵敏度,但仍需要专业技术人员和繁琐的实验室设备。即时诊断对结核病控制至关重要,因为它可以快速鉴定病原体以及结核菌株的耐药情况,这些耐药情况源于不同位点的单核苷酸多态性(SNP),从而能更准确地指示适当的治疗方法。金纳米颗粒已广泛应用于分子诊断平台。在此,我们描述了金纳米探针(寡核苷酸功能化金纳米颗粒)在一种非交联比色法中的应用,用于直接检测特定DNA靶标。由于金纳米颗粒具有显著的光学特性,该检测系统能够对比色法检测病原体,并有可能识别与结核病抗生素耐药性相关的几种单核苷酸多态性(SNP)。为了实现即时诊断,我们将这一策略应用于一种低成本的移动方案,该方案使用基于纸张的显色平台,并且通过智能手机设备将光谱特征转换为RGB数据。通过这种方式,可以在即时诊断时实现病原体的鉴定和耐药特征的表征。