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16S rRNA-functionalized multi-HCR concatemers in a signal amplification nanostructure for visual detection of Salmonella
Biotechnology and Applied Biochemistry ( IF 2.8 ) Pub Date : 2020-05-30 , DOI: 10.1002/bab.1962
Guijuan Qu 1 , Xiuwei Sun 1, 2 , Na Ying 2, 3 , Shengjun Bu 2 , Zhongyi Li 2 , Zhuo Hao 2 , Haibo Wu 4 , Jiayu Wan 2
Affiliation  

To prevent foodborne diseases and minimize their impacts, it is extremely important to develop a cost-effective and efficient bacterial detection assay for diagnostics, particularly in resource-poor settings. In this study, 16S rRNA from foodborne Salmonella was coupled with multiple HCR (hybridization chain reaction) concatemers and functionalized in a signal structure for lateral flow nucleic acid biosensor (LFNAB) detection. The 16S rRNA was incubated with two specific capture probes and multiple helper probes carrying the same initiator, to unwind its secondary structure and form an “initiators-on-a-string” complex. Through use of the initiators, each target 16S rRNA yielded multiple HCR concatemers tethered to numerous biotins, and numerous streptavidin-labeled gold nanoparticles were introduced on the LFNAB. The limit of detection was 53.65 CFU/mL for Salmonella. Notably, this method has high specificity and applicability for the detection of Salmonella in food and water samples.

中文翻译:

用于沙门氏菌视觉检测的信号放大纳米结构中的 16S rRNA 功能化多 HCR 串联体

为了预防食源性疾病并最大限度地减少其影响,开发一种具有成本效益且高效的细菌检测方法进行诊断极为重要,尤其是在资源匮乏的环境中。在本研究中,食源性沙门氏菌的16S rRNA与多个 HCR(杂交链反应)串联体结合,并在信号结构中功能化,用于侧流核酸生物传感器(LFNAB)检测。16S rRNA 与两个特定的捕获探针和多个携带相同起始子的辅助探针一起孵育,以解开其二级结构并形成“起始子串”复合物。通过使用引发剂,每个目标 16S rRNA 产生多个 HCR 多联体,连接到众多生物素,并且在 LFNAB 上引入了许多链霉亲和素标记的金纳米粒子。沙门氏菌的检测限为 53.65 CFU/mL 。值得注意的是,该方法对于食品和水样中沙门氏菌的检测具有很高的特异性和适用性。
更新日期:2020-05-30
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