Study on detection method of salmonella using aptamer- conjugated microbeads
-
摘要: 目的:研究基于核酸适体的微球芯片系统应用于沙门氏菌检测的可行性。方法:采用偶联有沙门氏菌适体的微球作为载体,以荧光作为检测信号建立微球芯片体系,并优化影响检测结果的各项因素。结果:通过此方法检测20份水产样品,6份检出沙门氏菌,检出率为30%。结论:建立的检测体系能实现对沙门氏菌的快速,准确的检测,且具有较高的特异性。Abstract: Objective: To investigate the feasibility of aptamer- conjugated microbeads for the detection of salmonella.Methods: Amino- terminal aptamer probes were covalently attached onto the surface of microbeads.Fluorescence from microbeads was detected as signal source, and parameters were optimized which may affect detection of the target.Results: Salmonella were detected in six of 20 aquatic product samples by this method, and the detection rate was 30%. Conclusion: The aptamer- conjugated microbeads can be used for high throughput and accurate detection of salmonella.
-
Keywords:
- salmonella /
- aptamer /
- microbeads /
- detection
-
[1] Collette F, Rachel S, Linda L G, et al.Molecular analysis of the rfb O antigen gene cluster of Salmonella enterica serogroup O:6 , 14 and development of a serogroup-specific PCR assay[J].Applied and Environmental Microbiology, 2003, 69 (10) :6099-6015.
[2] 范放, 洪小柳, 赵芳, 等.进口冷冻肉类沙门氏菌快速检验方法的研究[J].检验检疫学刊, 2009 (1) :45-48. [3] 刘晓霞.关于食品中沙门氏菌快速检测方法的新进展[J].化学工程与装备, 2009 (10) :133-134. [4] 江为民, 肖光明.关于我国水产品质量安全问题及应对措施[J].内陆水产, 2008 (1) :7-10. [5] 林涛, 兰敏, 黄伟, 等.荧光酶免疫分析仪-微生物鉴定仪在水产品沙门氏菌检测中的应用[J].食品与发酵工业, 2008, 34 (8) :133. [6] 王毳, 闰磊, 曾庆祝.沙门氏菌的检测技术与方法[J].现代食品科技, 2007, 23 (5) :82-85. [7] 王福厚, 霍贵成, 主德国, 等.食品中沙门氏菌检测方法的研究进展[J].中国乳品工业, 2009 (10) :38-41. [8] 余晓峰, 张萍, 宗凯, 等.免疫磁珠法检测脱水蒜制品中沙门氏菌[J].食品科学, 2012, 33 (24) :257-259. [9] 许学斌, 顾宝科, 金汇明, 等.免疫磁珠法检测食品中的沙门氏菌及分离菌株的耐药性[J].中国食品卫生杂志, 2006, 18 (3) :202-204. [10] 山珊, 牛瑞江, 等.免疫磁珠法富集沙门氏菌的优化及应用[J].食品工业科技, 2013, 34 (13) :153-156. [11] Sheng-Quan Jin, Bin-Cheng Yin, Bang-Ce Ye.Multiplexed Bead-Based Mesofluidic System for Detection of Food-Borne Pathogenic Bacteria[J].Applied and environmental microbiology, 2009, 75 (21) :6647-6654.
[12] Wolffs PF, Glencross K, Thibaudeau R, et al.Direct quantitation and detection of salmonella in biological samples without enrichment, using two-step filtration and real-time PCR[J].Appl.Environ.Microbiol, 2006, 72:3896-3900.
[13] Wang L., P.C.H.Li.Flexible microarray construction and fast DNA hybridization conducted on a microfluidic chip for greenhouse plant fungal pathogen detection[J].J.Agric.Food Chem, 2007, 55:10509-10516.
[14] R.Joshi, H.Janagama, H.P.Dwivedi.Selection, characterization, and application of DNA aptamers for the capture and detection of Salmonella enterica serovars[J].Mol.cell.Probes, 2009, 23:20-28.
[15] Gioi Dong Huy, Na Jin, Bin-Cheng Yin, et al.A novel separation and enrichment method of 17b-estradiol using aptamer-anchored microbeads[J].Bioprocess and Biosystens Engineering, 2011, 34 (2) :189-195.
[16] Ma X, Jiang Y, Jia F, et al.An aptamer-based electrochemical biosensor for the detection of salmonella[J].J Microbiol Methods, 2014, 3:94-98.
[17] Wenhe Wu, Jun Li, et al.Gold nanoparticle-based enzymelinked antibody-aptamer sanwich assay for detection of Salmonella typhimurium[J].ACS Appl mater interfaces, 2014, 6 (19) :16974-16981.
[18] Han Sheng, Bang-Ce Ye.Different strategies of covalent attachment of oligonucleotide probe onto glass beads and the hybridization properties[J].Appl Biochem Biotechnol, 2009, 152:54-65.
[19] Alexander WP, Lauren KW, Rosina MG.Hybridization of mismatched or partially matched DNA at surfaces[J].J AM CHEM SOC, 2002, 124:14601-14607.
[20] Nuzzo R G, Fusco F A, Allara D L.Spontuneously organized molecular assemblies (3) :preparation and properties of solution adsorbed monolayers of organic disulfides on gold surfaces[J].J Am Chem Soc, 1987, 109 (8) :2358-2361.
[21] Peng Zuo, Xiu Jun Li, Delfina C, et al.A PDMS/paper/glass hybrid microfluidic biochip integrated with aptamer-functionalized graphene oxide nano-biosensors for one-step multiplexed pathogen detection[J].Lab Chip, 2013, 13:3921-3928.
计量
- 文章访问数:
- HTML全文浏览量:
- PDF下载量:
下载:
