College of Environmental Science and Engineering, Qilu University of Technology (Shandong Academy of Science), Jinan, Shandong, 250353, China.
Department of Nursing, Shandong Medical College, No 5460 Erhuanan Road, Jinan, Shandong, 250002, China.
Chembiochem. 2024 Feb 1;25(3):e202300575. doi: 10.1002/cbic.202300575. Epub 2023 Dec 4.
Salmonella constitutes a prevalent alimentary pathogen, instigating zoonotic afflictions. Consequently, the prompt discernment of Salmonella in sustenance is of cardinal significance. Lateral flow assays utilizing colorimetric methodologies adequately fulfill the prerequisites of point-of-care diagnostics, however, their detection threshold remains elevated, generally permitting only qualitative discernment, an impediment to the preliminary screening of nascent pathogens. In response to this conundrum, we propose a lateral flow diagnostic predicated upon a streptavidin-biotin amplification system with recombinase polymerase amplification engineered for the expeditious and quantitative discernment of Salmonella enteritidis. Trace nucleic acids within a sample undergo exponential amplification via recombinase polymerase amplification to a level discernable, constituting the initial signal amplification. Subsequently, along the test line (T-line) of the lateral flow strip, the chromatic signal undergoes augmentation by securing a greater quantity of AuNPs through the magnification capacity of the streptavidin-biotin mechanism, affecting the second signal amplification. Quantitative results are procured via smartphone capture and transferred to computer software for precise calculation of the targeted quantity. The lateral flow strip exhibits a LOD at 19.41 CFU/mL for cultured S. enteritidis. The RSD of three varying concentrations were respectively 3.74 %, 5.96 %, and 4.25 %.
沙门氏菌构成了一种普遍的食源性病原体,引发动物传染病。因此,及时在食物中发现沙门氏菌至关重要。基于比色法的侧向流动分析能够充分满足即时诊断的要求,但它们的检测灵敏度仍然较高,通常只能进行定性检测,这对新生病原体的初步筛选构成了障碍。针对这一难题,我们提出了一种基于链霉亲和素-生物素放大系统的侧向流动诊断方法,并结合了重组酶聚合酶扩增技术,用于快速定量检测肠炎沙门氏菌。样本中的痕量核酸通过重组酶聚合酶扩增进行指数级扩增,达到可检测的水平,构成初始信号放大。随后,在侧向流动条的测试线(T 线)上,通过链霉亲和素-生物素放大机制捕获更多的 AuNPs,使显色信号得到增强,从而实现第二次信号放大。通过智能手机采集定量结果,并传输到计算机软件进行精确计算。该侧向流动条对培养的肠炎沙门氏菌的检测限为 19.41 CFU/mL。三个不同浓度的相对标准偏差分别为 3.74%、5.96%和 4.25%。