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利用双标记PCR产物的量子点检测副溶血性弧菌 被引量:4

Development of Quantum Dot by Double Labeling PCR Products for Detection of Vibrio parahemolyticus
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摘要 副溶血弧菌是一种能引起食源性疾病的重要病原菌,为了对该细菌进行检测,研制一种快速、灵敏、简便的量子点免疫层析试纸条。通过磁珠富集DNA后用双标记的特异性引物(上游引物5’用地高辛标记,下游引物5’用生物素标记)扩增不耐热溶血毒素基因(TLH),进行量子点免疫试纸条的检测。结果表明,荧光定量PCR检测磁珠(Si-MNPs)的捕获率在浓度102~106 CFU/m L均在50%以上,浓度为102 CFU/m L时达到最高捕获率(64%)。在纯培养物及模拟检测中试纸条的检测限均为4×100 CFU/m L,但在模拟检测中T线的显色强度弱于纯培养物的检测,说明鱼肉糜样本的复杂成分对检测灵敏度有一定的影响,但添加高浓度杂菌(阪崎肠杆菌、志贺氏菌)不影响该方法的特异性。方法具有特异性强、灵敏度高,并且操作简便、检测成本低的特点,2 h即可完成,为副溶血弧菌的快速检测提供了有效手段。 Vibrio parahemolyticus is an important pathogen which can cause food-borne illness.A rapid,sensitive and simple quantum dot immunity test strip was developed in order to detect bacteria.Genomic DNA was extracted with silica-coated magnetic nanoparticles,and a species-specific gene(thermolabile hemolysin,TLH) was amplified by PCR using digoxigenin/biotin-labeled primers,and used for immunochromatographic test strip(ICTS).The results showed that in the range of 10~2~10~6 CFU/mL,the extraction ratio of DNA of V parahemolyticus maintained above 50%,and the highest recovery(64%) by Si-MNP took place in 102 CFU/mL.The limit of detection for V parahemolyticus was 4 ×10° CFU/mL in pure culture and fish paste samples,respectively,which was not affected by those non-V parahemolyticus strains.However,detection color intensity is weaker in fish paste samples than in pure culture,indicating that the complex composition of fish meat sample has little influence s for the detection sensitivity.The method has high specificity,high sensitivity,and easy to operate,low detection cost,accomplished in two hour,and provides an effective means for rapid detection of V parahaemolyticus.
出处 《食品工业》 北大核心 2015年第6期283-287,共5页 The Food Industry
基金 国家自然科学基金项目(31000048,31170091) 食品科学与技术国家重点实验室目标导向课题(SKLF-MB-201002) 自由探索课题(SKLF-TS-200916) 赣鄱英才555工程人才计划(2012年)
关键词 副溶血弧菌 量子点 免疫层析试纸条 Vibrio parahemolyticus quantum dot immunochromatographic test
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参考文献23

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二级参考文献108

共引文献244

同被引文献102

  • 1刘金华,刘韬,孟日增,聂丹丹,刘阳,薛力刚.利用光纤倏逝波生物传感器检测食品中大肠杆菌O157:H7[J].食品安全质量检测学报,2014,5(4):1142-1146. 被引量:6
  • 2刘敏,刘兆阅,吕强,袁航,马岚,李景虹,白玉白,李铁津.Functionalized Fluorescein-doped SiO2 Nanoparticles for Immunochromatographic Assay[J].Chinese Journal of Chemistry,2005,23(7):875-880. 被引量:1
  • 3李世敏.美国食源性疾病监测预警系统及其特点[J].中国卫生监督杂志,2005,12(6):434-437. 被引量:21
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