食品科学 ›› 2011, Vol. 32 ›› Issue (12): 183-187.doi: 10.7506/spkx1002-6630-201112040

• 分析检测 • 上一篇    下一篇

应用PCR技术快速检测腐烂苹果中扩展青霉

何鸿举,焦凌霞,樊明涛*,张建兵,吕丽娟,刘晓娇,李亚菲   

  1. 1. 西北农林科技大学食品科学与工程学院
    2.河南科技学院食品学院 3.陕西出入境检验检疫局国际旅行卫生保健中心
  • 出版日期:2011-06-25 发布日期:2011-06-10
  • 基金资助:
    教育部博士点基金项目(200807120017);陕西省科技攻关项目(2007K01-12)

Rapid Detection of Penicillium expansum in Rotten Apples by Polymerase Chain Reaction

HE Hong-ju1,JIAO Ling-xia2,FAN Ming-tao1,*,ZHANG Jian-bing3,LU Li-juan1,LIU Xiao-jiao1,LI Ya-fei1   

  1. 1. College of Food Science and Engineering, Northwest A & F University, Yangling 712100, China; 2. College of Food Science, Henan Institute of Science and Technology, Xinxiang 453003, China; 3. Shaanxi International Travel Healthcare Center, Shaanxi Entry-Exit Inspection and Quarantine Bureau, Xi’an 710068, China
  • Online:2011-06-25 Published:2011-06-10

摘要: 利用扩展青霉的多聚半乳糖醛酸酶基因序列,设计一对特异性引物,通过聚合酶链式反应(polymerase chain reaction,PCR)反应快速检测腐烂苹果中的扩展青霉。反应的特异性和敏感性以及反应的最适条件等试验的结果表明:该引物具有很高的特异性和灵敏性,只扩增扩展青霉DNA,而不扩增其他真菌DNA;菌体的检测限为1.34×103个孢子/mL,DNA的检测限为2.40×10-2μg/mL;反应的最适退火温度范围为54~59℃,最适模板浓度范围为2.40~5.28μmol/L。该方法具有简单、快速、特异性好和灵敏性高等特点,可为快速检测腐烂苹果中扩展青霉的实际应用提供借鉴。

关键词: 模板, 引物, 特异性, PCR检测, 扩展青霉

Abstract: In this study, a pair of specific primers was designed according to the polygalacturonase gene of Penicillium expansum to detect Penicillium expansum in rotten apples by polymerase chain reaction (PCR). The results showed that the specificity and sensitivity of the design primer pair were high and only Penicillium expansum DNAs were amplified. The detection limits of Penicillium expansum and DNA were 1.34 × 103 spores/mL and 2.40 × 10-2μg/mL, respectively. The optimal reaction condition was obtained as follows: anneal temperature of 54-59 ℃ and template concentration of 2.40-5.28μg/mL. Owing to its simplicity, rapidity and high specificity and sensitivity, this assay can be used for practical applications.

Key words: PCR(polymerase chain reaction)detection, penicillium expansum, template, primers, specificity

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