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基于比色法和微流控芯片的搅拌棒吸附萃取双模适体传感器同时检测多种食源性病原菌
Microchimica Acta
(
IF
5.7
)
Pub Date : 2021-07-06
, DOI:
10.1007/s00604-021-04902-1
Ming Wang
1
,
Jin Zeng
1
,
Jiaqi Wang
1
,
Xiao Wang
1
,
Ye Wang
2
,
Ning Gan
1
Affiliation
- Faculty of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, China.
- Zhejiang Pharmaceutical College, Ningbo, 315211, China.
已开发出一种使用比色法和微流控芯片 (MC) 以及搅拌棒吸附萃取 (SBSE) 的双模式适体传感器,用于首次鉴定受副溶血性弧菌(VP) 和鼠伤寒沙门氏菌污染的样品(ST),然后在正样本中精确确定它们。为此,预先制备了对应于不同细菌的适体-链霉亲和素编码探针(Apt-SAEs)。然后,用 4-巯基苯基硼酸 (MPBA) 改性的搅拌棒与 Apt-SAE 探针一起提取细菌。适体与靶标的结合事件触发了含有Apt-SAE、VP或ST的两个夹心结构的形成,细菌浓度可在15分钟内富集1000倍,避免了长时间的富集过程。最后,将搅拌棒浸入 3,3',5,5'-四甲基联苯胺 (TMB)-H 2 O 2显色解决方案。肉眼可以观察颜色来判断分析物是否存在。无色样品判断为阴性。对于阳性样品,对应于不同细菌的吸附编码探针将从搅拌棒上洗脱下来,并由 MC 快速分析。在优化的条件下,比色法可以观察到 100 CFU/mL 的 VP 或 ST 或两者,而 MC 可以检测到 35 CFU/mL(S/ N = 3)。该方法对食源性病原菌的现场筛查和多重检测具有重要的应用价值。
图形概要
"点击查看英文标题和摘要"
Dual-mode aptasensor for simultaneous detection of multiple food-borne pathogenic bacteria based on colorimetry and microfluidic chip using stir bar sorptive extraction
A dual-mode aptasensor using colorimetry and microfluidic chip (MC) together with stir bar sorptive extraction (SBSE) has been developed for firstly qualifying samples contaminated with Vibrio parahaemolyticus (V.P) and Salmonella typhimurium (S.T), then precisely determine both of them in positive samples. For this purpose, the aptamer-streptavidin encoded probes (Apt-SAEs) corresponding to different bacteria were prepared in advance. Then, a stir bar modified with 4-mercaptophenylboronic acid (MPBA) was made to extract bacteria together with Apt-SAE probes. The binding event of aptamer and target triggered the formation of two sandwich structures containing Apt-SAE, V.P or S.T. The concentration of bacteria could be enriched by 1000 times within 15 min to avoid long-time enrichment process. Finally, the stir bar was immersed in the 3,3′,5,5′-Tetramethylbenzidine (TMB)-H2O2 solution for color development. The color could be observed by naked eyes to judge whether the analytes were present. The colorless samples were judged to be negative. For the positive samples, the adsorbed encoded probes corresponding to different bacteria would be eluted from the stir bar and rapidly analyzed by the MC. Under the optimized conditions, 100 CFU/mL of V.P or S.T or both of them could be observed by colorimetry and 35 CFU/mL of them could be detected (S/N = 3) by the MC. The assay has significant application value for on-site screening and multiple detection of food-borne pathogenic bacteria.
Graphical abstract
更新日期:2021-07-07