| 用于检测仔猪致病菌的23S rRNA基因芯片的研制 |
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| 引用本文： | 伍诚意,曹峰子,梁之昶,章振华,杨兵,周宏专,季海峰,陈小玲.用于检测仔猪致病菌的23S rRNA基因芯片的研制[J].中国兽医科学,2010,40(3). |
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| 作者姓名： | 伍诚意 曹峰子 梁之昶 章振华 杨兵 周宏专 季海峰 陈小玲 |
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| 作者单位： | 伍诚意(北京市农林科学院,畜牧兽医研究所,北京,100097;江西生物科技职业学院,江西,南昌,330200);曹峰子(北京市农林科学院,畜牧兽医研究所,北京,100097;河北农业大学,动物科技学院,河北,保定,071000);梁之昶(北京市农林科学院,畜牧兽医研究所,北京,100097;北京华大吉比爱生物技术有限公司,北京,101300);章振华,杨兵,季海峰,陈小玲(北京市农林科学院,畜牧兽医研究所,北京,100097);周宏专(北京市农林科学院,畜牧兽医研究所,北京,100097;首都师范大学,生命科学院,北京,100037) |
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| 摘 要： | 以副猪嗜血杆菌、胸膜肺炎放线杆菌、多杀性巴氏杆菌、大肠杆菌、肠道沙门氏菌、猪链球菌等12种仔猪致病菌为目标细菌,23SrRNA基因为靶基因,从GenBank中下载其23SrDNA全序列,在其变异区设计特异性探针,在保守区设计通用引物。通过序列分析、PCR靶标与探针杂交试验,建立了有13条寡核苷酸探针和2对通用引物的仔猪常见致病菌基因芯片检测方法。以参考菌株粗提基因组DNA的10倍系列稀释模板做PCR,扩增产物与探针杂交,检出芯片的灵敏度为100fg。对33个参考菌株用芯片检测,结果有1例不正确,只鉴定到属;在61个野外分离株中,55株的芯片检测结果与生化或测序结果一致,有6例不一致,符合率为90%。初步研究证明,该检测方法能快速、有效地鉴定多种仔猪常见致病菌,但要区分某些细菌的致病株与非致病株,还要检测毒力/毒素基因。
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| 关 键 词： | 仔猪 致病菌 23S rRNA基因 芯片 寡核苷酸探针 |
Development of 23S rRNA gene-based microarray for detection of pathogenic bacteria in piglets |
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| WU Cheng-yi,CAO Feng-zi ,LIANG Zhi-chang,ZHANG Zhen-hua,YANG Bing,ZHOU Hong-zhuan,JI Hai-feng,CHEN Xiao-ling.Development of 23S rRNA gene-based microarray for detection of pathogenic bacteria in piglets[J].Veterinary Science in China,2010,40(3). |
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| Authors: | WU Cheng-yi CAO Feng-zi LIANG Zhi-chang ZHANG Zhen-hua YANG Bing ZHOU Hong-zhuan JI Hai-feng CHEN Xiao-ling |
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| Institution: | WU Cheng-yi1,2,CAO Feng-zi 1,3,LIANG Zhi-chang1,4,ZHANG Zhen-hua1,YANG Bing1,ZHOU Hong-zhuan1,5,JI Hai-feng1,CHEN Xiao-ling1(1.Institute of Animal Science , Veterinary Medicine,Beijing Municipal Academy of Agriculture , Forestry,Beijing 100097,China,2.Vocational College of Bio-science , Technology,Nanchang 330200,3.College of Animal Science , Technology,Hebei Agricultural University,Baoding 071000,4.Beijing BGI-GBI Biotech Co.,Ltd.,Beijing 101300,5.College of Life Sciences,Capital ... |
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| Abstract: | To develop a DNA microarray for rapid detection of pathogenic bacte-ria in piglets,12 bacterial species such as Haemophilus parasuis,Actinobacillus pleuropneumonia,Pasteurella multocida,Escherichia coli,Salmonella enterica and Streptococcus suis were chosen as the targets and 23S rDNA sequences of the bacteria were downloaded from the GenBank.The variable and conserved regions of the sequences were allocated for designing probes and primers.Based on sequence analysis and hybridization tests of PCR amplicons... |
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| Keywords: | piglet pathogenic bacteria 23 S rRNA gene microarray oligonucleotide probe |
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