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柑桔溃疡病菌PCR检测方法(引物)筛选

Screening of primers for PCR detection of Xanthomonas citri pv.citri

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为准确检测柑桔溃疡病菌,减少检测过程中假阳性和假阴性结果,进而为农业执法部门和口岸一线检疫鉴定提供参考,采用已报道的14对常规PCR和实时荧光PCR引物/探针,对来自不同国家和地区的22株柑桔溃疡病菌及9株黄单胞菌属其他菌株进行了特异性测试和灵敏度比对分析,对优选引物采用9份具柑桔溃疡病症状的样品进行检测验证.结果表明,引物对JYF5(XccF05)/JYR5(XccR05)和Xac01/Xac02的检测特异性高,对柑桔溃疡病菌DNA的检测灵敏度均为3 pg/μL;其他12对引物/探针会出现不同程度上的假阳性或假阴性,其中,XAcF/XAcR对来自泰国的酸橙样品和阳性菌株进行检测均为假阴性.推荐JYF5(XccF05)/JYR5(XccR05)和Xac01/Xac02这2对引物用于柑桔溃疡病菌的检测.
In order to accurately detect Xanthomonas citri pv.citri,reduce false positive and false negative detection results,and provide reference for quarantine identification for agricultural law enforcement depart-ments and ports,14 pairs of conventional PCR and real-time fluorescence PCR primers and probes were used for comparison analysis of the detection specificity and sensitivity of 22 strains of X.citr i pv.citri and 9 other strains of Xanthomonas from different countries and regions.The selected primers were tested and verified by 9 samples with suspected symptoms of citrus canker.The results showed that the primer pairs of JYF5(XccF05)/JYR5(XccR05)and Xac01/Xac02 had higher specificity,and the detection sensitivity of both to X.citri pv.citri DNA were 3 pg/μL.The other 12 pairs of primers and probes showed different degree of false positive or false negative.Among them,XAcF/XAcR was false negative for Citrus aurantifolia samples and positive strains from Thailand.Based on the above results,JYF5(XccF05)/JYR5(XccR05)and Xac01/Xac02 were recommended for the detection of X.citri pv.citri.

Xanthomonas citri pv.citriprimerconventional PCRreal-time PCR

滕少娜、孙涛、周林、孔德英、姚廷山、李敏、易建平

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重庆海关技术中心,重庆,400020

西南大学柑桔研究所,重庆,400712

福州海关技术中心,福州,350001

上海海关动植物与食品检验检疫技术中心,上海,200002

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柑桔溃疡病菌 引物 常规PCR 实时荧光PCR

2024

中国南方果树
中国农业科学院柑桔研究所

中国南方果树

CSTPCD北大核心
影响因子:0.527
ISSN:1007-1431
年,卷(期):2024.53(6)