应用PCR技术快速检测腐烂苹果中扩展青霉菌  

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

作　　者：何鸿举[1] 焦凌霞[2] 樊明涛[1] 张建兵 吕丽娟[1] 刘晓娇[1] 李亚菲[1] 

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100 [2]河南科技学院食品学院,河南新乡453003 [3]陕西出入境检验检疫局国际旅行卫生保健中心,陕西西安710068

出　　处：《食品科学》2011年第12期183-187,共5页Food Science

基　　金：教育部博士点基金项目(200807120017);陕西省科技攻关项目(2007K01-12)

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 chainreaction,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。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.

关 键 词：聚合酶链式反应(PCR)检测 扩展青霉 DNA模板 引物特异性 

分 类 号：Q939.92[生物学—微生物学]