全文下载：2012020164
石琰璟1 ， 王建广1,2
（1. 青岛科技大学 化工学院， 山东 青岛 266042; 2. 山东省出入境检验检疫局, 山东 青岛 266022)
摘要: 采用自行建立和优化的PCR-核酸薄膜层析检测体系，对创伤弧菌(Vibrio vulnificus)进行检测。采用创伤弧菌的vvhA基因为目的片段设计特异性引物，建成可快速检测创伤弧菌的PCR-核酸薄膜层析检测法，并进行了特异性和灵敏度试验。结果表明，所建立起的检测法灵敏度为3.6×102 cfu·mL-1，比PCR-琼脂糖凝胶电泳检测方法高一个数量级。创伤弧菌的PCR-核酸薄膜层析检测方法具有较高灵敏度和良好特异性，操作简单，检测速度快，同时又保护了操作人员的身体健康，具有广阔的推广前景。
关键词: 创伤弧菌； 检测； PCR； 薄膜层析
中图分类号: R 155.3文献标志码： A
Establishment of a PCR-Nucleic Acid Thin Layer Chromatography Assay to Detect Vibrio VulnificusSHI Yan-jing1, WANG Jian-guang2
(1. College of Chemical Engineering，Qingdao University of Science and Technology，Qingdao 266042，China;
2. Shandong Entry-exit Inspection and Quarantine Bureau,Qingdao 266022,China）
Abstract: A new method was established based on PCR-Nucleic Acid Thin Layer Chromatography assay to detect Vibrio vulnificus. A set of highly specific primers were synthesized according to the vvhA gene of Vibrio vulnificus so as to the establish PCR-Nucleic Acid Thin Layer Chromatography Assay. Specificity and sensitivity of the method were tested.The results showed that the sensitivity of PCR-Nucleic Acid Thin Layer Chromatography of Assay was 3.6×102 cfu·mL-1 with 10 times higher than that of PCR-Agarose Gel Electrophoresis Assay, and the specificity is also satisfied. This method showed its wide perspective and practicability by its simpilicity，high specficity and sensitivty on detecting Vibrio vulnificus.
Key words: Vibrio vulnificus； detection； PCR； thin layer chromatography
收稿日期：2011-06-29
作者简介： 石琰璟(1974—)，女，副教授.
