[1]李伟杰,赵 耘*,杜昕波,等.产毒素多杀性巴氏杆菌菌落双重PCR检测方法的建立[J].中国预防兽医学报,2010,(04):298-300.
 LI Wei-jie,ZHAO Yun*,DU Xin-bo,et al.Detection of toxigenic Pasteurella multocida with a colony diplex PCR assay[J].Chinese journal of preventive veterinary medicine,2010,(04):298-300.
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产毒素多杀性巴氏杆菌菌落双重PCR检测方法的建立
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2010年04期
页码:
298-300
栏目:
研究简报
出版日期:
2010-04-01

文章信息/Info

Title:
Detection of toxigenic Pasteurella multocida with a colony diplex PCR assay
文章编号:
1008-0589(2010)04-0298-03
作者:
李伟杰赵 耘*杜昕波康 凯陈 敏
中国兽医药品监察所,北京 100081
Author(s):
LI Wei-jie ZHAO Yun* DU Xin-bo KANG Kai CHEN Min
China Institute of Veterinary Drug Control, Beijing 100081, China
关键词:
产毒素多杀性巴氏杆菌双重PCRKMT1基因toxA基因
Keywords:
toxigenic Pasteurella multocida diplex PCR KMT1 gene toxA gene
分类号:
S852.61
文献标志码:
A
摘要:
为建立快速特异的PCR方法以及同时检测并区分产毒素与非产毒素多杀性巴氏杆菌,本研究根据GenBank登录的多杀性巴氏杆菌KMT1基因和toxA毒素基因序列,设计合成了2对特异引物。特异性试验表明产毒素多杀性巴氏杆菌C51-6扩增出了460 bp和1 854 bp的2条目的片段,而不产毒素多杀性巴氏杆菌、大肠埃希菌、胸膜肺炎放线杆菌、猪链球菌、支气管败血波氏杆菌、副猪嗜血杆菌和鸡白痢沙门菌的扩增均为阴性;敏感性试验表明该PCR方法能从含450 CFU的菌液中扩增出相应的目的片段。同时用豚鼠皮肤坏死试验和小鼠致死试验对该PCR方法进行了验证。
Abstract:
A duplex PCR assay was developed using two sets of specific primers derived from the KMT1 gene and toxA gene of Pasteurella multocida in order to differentiate toxigenic and nontoxigenic P. multocida at the same time. The assay could specifically amplify both 460 bp and 1 854 bp DNA products from thee toxigenic P. multocida C51-6, but not from nontoxigenic P. multocida, E. coli, Actinobacillus pleuropneumoniae, Streptococcus suis, Bordetella bronchiseptica, Haemophilus parasuis and Salmonella enterica subsp. It is highly sensitive and could detect as little as 450 CFU bacteria. The assay was verifiable by Guinea pig skin test and mouse lethal test.

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备注/Memo

备注/Memo:
收稿日期:2009-09-09
基金项目:国家科技支撑计划(2006BAD06A11)
作者简介:李伟杰(1979-),男,山东莱州人,助理研究员,主要从事兽医微生物资源开发和利用的研究.
更新日期/Last Update: 2010-05-19