文献类型：期刊文献

中文题名：美国进境樱桃果实中梨火疫病菌的检测

英文题名：Detection of Erwinia amylovora in cherry fruits imported from USA by PCR

作者：尚琳琳[1];周国梁[2];仇书红[2];陈仲兵[2];徐殿胜[1];易建平[2]

机构：[1]华东理工大学生物工程学院,上海200237;[2]上海出入境检验检疫局,上海200135

年份：2010

卷号：37

期号：5

起止页码：441

中文期刊名：植物保护学报

外文期刊名：Journal of Plant Protection

收录：CSTPCD;;北大核心:【北大核心2008】；CSCD:【CSCD2011_2012】；

基金：上海出入境检验检疫局科研项目(HK069-2010);上海市科委项目(09DZ0580201)

语种：中文

中文关键词：梨火疫病菌;樱桃果实;常规PCR;套式PCR;实时荧光PCR

外文关键词：Erwinia amylovora; cherry fruits; conventional PCR; nested PCR; real-time PCR

摘要：为了筛选快速、灵敏的梨火疫病菌检测方法,利用常规PCR、套式PCR和实时荧光PCR方法分别对美国进境的326批樱桃果实中梨火疫病菌进行检测。结果显示,3种PCR方法的检出率不同,不同引物或探针的检出率也存在差异。在常规PCR中,引物Ams3/Ams4c、P29A/P29B和PEANT1/PEANT2的检出率分别为35.28%、24.85%和16.87%;单管套式PCR和套式PCR的检出率分别为23.01%和50.61%;4种实时荧光PCR的检出率分别为17.48%(探针PA)、32.21%(探针Ams)、29.14%(探针ITS)和23.93%(SYBR GreenⅠ)。在所有试验方法中由引物P29A/P29B和PEANT1/PEANT2组成的套式PCR的检出率最高。检测结果证实了进境樱桃果实中存在梨火疫病菌DNA,套式PCR和常规PCR(引物Ams3/4c)可用于进境樱桃样品中梨火疫病菌的常规检测。
In order to find out the fast and efficient detection method for Erwinia amylovora,three methods including the conventional PCR,nested PCR and real-time PCR were evaluated to detect E.amylovora in cherry fruits imported from USA.The results of detection with 326 samples of cherry fruits showed that the primers Ams3/Ams4c,P29A/P29B and PEANT1/PEANT2 in the conventional PCR achieved 35.28%,24.85% and 16.87% of positive samples,respectively.The percent of positive samples for the nested PCR in single closed tube was 23.01% and 50.61% for conventional nested PCR.The prob PA,prob Ams,prob ITS and SYBR GreenⅠin real-time PCR detected 17.48%,32.21%,29.14% and 23.93% of positive percent,respectively.The nested PCR combined with primers P29A/P29B and PEANT1/PEANT2 showed the highest positive samples numbers with 50.61%.The results of detection and sequence analysis approved the existence of E.amylovora DNA in cherry fruits imported from USA.The nested PCR and the primers Ams3/4c in the conventional PCR are proposed for routine use in quarantine detection of E.amylovora in cherry fruits.