中文摘要：本发明公开了一种猕猴桃细菌性溃疡病菌的检测方法，包括：将细菌ITS通用引物对所有猕猴桃溃疡病菌菌株扩增测序；对其进行ITS序列进行多重同源性比较分析，设计猕猴桃溃疡病菌特异引物和探针；对猕猴桃溃疡病菌特异引物和探针进行可用性检测、特异性检测、以菌液为模板的灵敏度检测、以DNA为模板的灵敏度检测、以接种猕猴桃枝条为对象的荧光PCR检测和以田间发病样品为对象的荧光PCR检测；直至获得实时荧光PCR引物和实时荧光PCR探针。本实发明可以有效地防止检测过程中的污染和假阳性结果，得到实时荧光PCR引物和探针，建立猕猴桃溃疡病菌实时定量荧光PCR检测体系，实现猕猴桃溃疡病菌的快速、特异、灵敏、定量检测。
 
外文摘要：NOVELTY - Detecting pathogen of kiwifruit bacterial ulcer involves using standard primers having ITS region of 16S-23S recombinant DNA of Kiwifruit ulcer pathogens which are amplified by polymerase chain reaction and sequenced. The biology software clastal X is used to compare the sequences after sequencing and the homologous ITS sequences of the Pseudomonas syringae in the GenBank database to detect the mutations of kiwifruit ulcer pathogens.
 
USE - The method enables to detect pathogen of kiwifruit bacterial ulcer, which shows fast, specific and sensitive quantitative detection.
 
DETAILED DESCRIPTION - Detecting pathogen of kiwifruit bacterial ulcer involves using standard primers having ITS region of 16S-23S recombinant DNA of Kiwifruit ulcer pathogens which are amplified by polymerase chain reaction and sequenced. The biology software clastal X is used to compare the sequences after sequencing and the homologous ITS sequences of the Pseudomonas syringae in the GenBank database to detect the mutations of kiwifruit ulcer pathogens. The stability points of kiwi fruit ulcer pathogens, primers and probe is designed using primer Express software for obtaining kiwi ulcer pathogen specific primers and kiwi fruit ulcer pathogens. The sensitivity of kiwifruit ulcer pathogen is tested. The sensitivity test with bacteriophage as template involves selecting DNA as template, and kiwi fruit fluorescent polymerase chain reaction is used to detect the target. The primers and probe designing using software Prmier express and the sensitivity detection step is r!
 epeated until a real-time fluorescent polymerase chain reaction primer and a real-time fluorescent polymerase chain reaction probe for the detection of kiwifruit bacterial ulcer pathogens are obtained.
 
主权项：
申请号：CN201611158447.7
公开/公告号：CN106399578A
申请日：2016-12-15
公开/公告日：2017-02-15
申请/专利权人：安发（福建）生物科技有限公司
发明/设计人：高益槐;郑春源;戴金玉;唐文波
分类号：C12Q1/68;C12Q1/04;C12R1/38
主分类号：C12Q1/68
法律状态：实质审查
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