Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology , Wuhan University , Wuhan 430072 , People's Republic of China.
Wuhan Academy of Agricultural Sciences , Wuhan 430072 , People's Republic of China.
Anal Chem. 2019 Jan 2;91(1):1178-1184. doi: 10.1021/acs.analchem.8b05154. Epub 2018 Dec 18.
Rapid and sensitive foodborne pathogen detection assay, which can be applied in multiple fields, is essential to timely diagnosis. Herein, we proposed a multisignal readout lateral flow immunoassay for Salmonella typhimurium ( S. typhi) detection. The assay employs colorimetric-fluorescent-magnetic nanospheres (CFMNs) as labels, which possess multifunctional target separation and enrichment, multisignal readout, and two formats of quantitation. The assay for S. typhi detection involves magnetic separation and chromatography. First, the S. typhi were separated and enriched from matrix by antibody labeled CFMNs, and then the S. typhi-containing suspension is added onto the sample pad to flow up the test strip. The introduction of magnetic separation enhances anti-interference ability and 10-fold sensitivity, making the assay possible for practical application. The assay has realized naked eye detection of 1.88 × 10 CFU/mL S. typhi, and 3.75 × 10 CFU/mL S. typhi can be detected with a magnetic assay reader, which is 2-4 orders of magnitude lower than other label-based LFIAs, with a quantitation range of 1.88 × 10 to 1.88 × 10 CFU/mL by measuring the fluorescence intensity and magnetic signal. Moreover, the successful detection of S. typhi in complex matrix (tap water, milk, fetal bovine serum, and whole blood) indicated its potential application in real samples.
用于食源性致病菌快速、灵敏检测的方法,在多个领域都具有重要意义,因为它可以实现及时诊断。在此,我们提出了一种基于多信号读取的侧向流动免疫分析方法,用于检测鼠伤寒沙门氏菌(S. typhi)。该分析方法采用比色-荧光-磁性纳米球(CFMNs)作为标记物,具有多功能的目标分离和富集、多信号读取以及两种定量格式。用于检测 S. typhi 的分析方法涉及到磁分离和色谱技术。首先,通过抗体标记的 CFMNs 将 S. typhi 从基质中分离和富集,然后将含有 S. typhi 的悬浮液添加到样品垫上,使其在测试条上流动。磁分离的引入增强了抗干扰能力,灵敏度提高了 10 倍,使该分析方法能够实际应用。该分析方法实现了对 1.88×10 CFU/mL S. typhi 的肉眼检测,使用磁性分析读取器可以检测到 3.75×10 CFU/mL 的 S. typhi,比其他基于标记的 LFIAs 低 2-4 个数量级,通过测量荧光强度和磁信号,其定量范围为 1.88×10 至 1.88×10 CFU/mL。此外,在复杂基质(自来水、牛奶、胎牛血清和全血)中成功检测到 S. typhi,表明其在实际样品中的潜在应用。