Sensitive colorimetric immunoassay of Vibrio parahaemolyticus based on specific nonapeptide probe screening from a phage display library conjugated with MnO2 nanosheets with peroxidase-like activity
Sensitive colorimetric immunoassay of Vibrio parahaemolyticus based on specific nonapeptide probe screening from a phage display library conjugated with MnO2 nanosheets with peroxidase-like activity
复制标题
基于从与具有过氧化物酶样活性的 MnO2 纳米片缀合的噬菌体展示文库中筛选的特异性九肽探针，对副溶血弧菌进行灵敏的比色免疫分析
DOI：
10.1039/c7nr06633c
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发表时间：
2018-02-14
期刊：
影响因子：
6.7
通讯作者：
Liu, Aihua
中科院分区：
文献类型：
--
作者：
Liu, Pei;Han, Lei;Liu, Aihua
由于病原体对公众健康的有害影响，病原体检测继续受到极大的关注。本研究从F8/9景观噬菌体文库中以高通量的方法从VQTVQIGSD噬菌体中分离到特异性的非肽融合蛋白pVIII(pVIII融合蛋白)。所制备的副溶血性弧菌特异性识别元件比抗体更便宜，更容易获得。在此基础上，以pVIII融合蛋白为捕获探针，以蛋白模板二氧化锰纳米片(NSS)为信号探针，建立了检测副溶血性弧菌的高灵敏度比色免疫分析方法。在靶细菌副溶血性弧菌存在的情况下，pVIII融合-V的夹心型复合体。副溶血性弧菌-MnO2 NS@pVIII融合是通过对pVIII融合和副溶血性弧菌的特异性识别而形成的。信号探针(MnO2-NSS)能催化3，3‘，5，5’-四甲基联苯胺与H_2O_2发生显色反应。所建立的副溶血性弧菌检测方法具有检测范围宽(20-10(4)个菌落形成单位(CFU)m L(-1))、低检测限(15 cfu m L(-1))、良好的选择性和高可靠性，在海洋微生物检测和控制方面具有潜在的应用前景。
Pathogen detection continues to receive significant attention due to the harmful effects of pathogens on public health. Herein, specific nonapeptide-fusion proteins pVIII (pVIII fusion) were isolated from phage VQTVQIGSD (designated by the sequence of a fused foreign peptide), which was specifically screened from the f8/9 landscape phage library against Vibrio parahaemolyticus (V. parahaemolyticus) in a highthroughput way. The as-prepared V. parahaemolyticus-specific recognition element is cheaper and more available than antibodies. Further, a highly sensitive colorimetric immunoassay for V. parahaemolyticus was established using pVIII fusion as capture probes coupled with protein-templated MnO2 nanosheets (NSs) as signal probes. In the presence of a target bacterium, V. parahaemolyticus, a sandwich-type complex of pVIII fusion-V. parahaemolyticus-MnO2 NS@pVIII fusion was formed through specific recognition of pVIII fusion and V. parahaemolyticus. The signal probes (MnO2 NSs) could catalyze the reaction of 3,3', 5,5'-tetramethylbenzidine and H2O2 to generate a colorimetric change. The proposed V. parahaemolyticus detection method demonstrated a wide detection range (20-10(4) colony-forming units (CFU) mL(-1)), low limit of detection (15 CFU mL(-1)), excellent selectivity, and high reliability for real marine samples, showing potential application in marine microbiological detection and control.