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副猪嗜血杆菌aroA基因的克隆与序列分析及2种三重PCR检测方法的建立 被引量:1

Isolation and Identification of Haemophilus parasuis,Cloning and Sequence Analysis of aroA Gene and Institution of Two Program of Triplex PCR Assay
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摘要 本试验对分离的5株副猪嗜血杆菌进行了"卫星"生长试验、生化鉴定和PCR检测。通过对aroA基因序列进行分析,发现这5株菌同标准菌株相比,其核苷酸序列相似性为96.3%~99.8%,氨基酸序列相似性为98.2%~99.5%。此外,还建立了分别针对副猪嗜血杆菌(Hps)、猪伪狂犬病病毒(PRV)、猪细小病毒(PPV)及猪圆环病毒2型(PCV2)、猪细小病毒、猪伪狂犬病病毒的三重PCR检测方法;敏感性试验表明,三重PCR最低能检出Hps、PPV、PRV、PCV2的DNA分别为12、16、7.6和6.3 pg。 In this study,5 strains of Haemophilus parasuis(Hps)were identified by 'satellite' growing test,biochemical and PCR assay.The results showed that aroA gene of the isolated Hps had a high degree of similarity with classical culture strain in the sequence of nucleotide and amino acid sequence,which was between 96.3% to 99.8% and 98.2% to 99.5% respectively.We implemented two program of triplex PCR assay,one group was used to detect simultaneously infections of Hps,PCV2 and PPV,the other group was developed to detect the PCV2,PPV and PRV.The sensitivity tests indicated that the triplex PCR assay may be used for detecting Hps,PPV,PRV and PCV2 of 105 dilute strength;the DNA minimun detectable quantity of Hps,PPV,PRV,PCV2 was 12,16,7.6 and 6.3 pg respectively.
出处 《中国畜牧兽医》 CAS 北大核心 2012年第8期10-15,共6页 China Animal Husbandry & Veterinary Medicine
关键词 副猪嗜血杆菌 三重PCR aroA基因 Haemophilus parasuis triplex PCR assay aroA gene
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  • 1Oliveira S, Pijoan C. Haemophilus parasuis : new trends on diagnosis, epidemiology and control [J]. Vet Microbiol, 2004(99) :1-12.
  • 2Simone Oliveira, Lucina Galina, Carlos Pijoan. Develop ment of a PCR test to diagnose Haemophilus parasuis infections[J]. J Veterinary Diagnostic Investigation, 2001 (13) :495--501.
  • 3Фystein Angen, Simone Oliveira, Peter Ahrens, et al. Development of an improved species specific PCR test for detection of Haemophilus parasuis [J]. Veterinary Mi- crobiology, 2007,119 (2/4) : 266-276.
  • 4Jung K, Ha Y, Kim S H, et al. Development of poly-merase chain reaction and comparison with in situ hybrid- ization for the detection of Haemophilus parasuis in for- malin-fixed, paraffin-embedded tissues [J]. J Vet Med Sci, 2004,66 (7) : 841 -- 845.
  • 5Frandoloso R, Martinez-Martinez S, Rodriguez-Ferri E F,et al. Comparison of real-time PCR and culture isola tion in colostrum-deprived pigs immunized and challenged with Haemophilus parasuis [J]. Lett Appl Microbiol, 2012,54(2) : 149 - 152.
  • 6魏晓嫒.副猪嗜血杆菌环介导等温扩增快速检测方法的建立[M].保定:河北农业大学,2013.
  • 7李军,谢宇舟,禤雄标,陈泽祥,杨威,马春霞,胡帅,彭昊,许力干,谢永平,潘艳.副猪嗜血杆菌实时荧光PCR快速检测方法的建立和应用[J].安徽农业科学,2011,39(6):3607-3609. 被引量:14
  • 8陈善真,李春玲,贾爱卿,王贵平.副猪嗜血杆菌OMP5基因的克隆、表达及间接ELISA检测方法的建立[J].中国农业科学,2011,44(14):3036-3044. 被引量:22
  • 9朱必凤,杨旭夫,彭凌,韦昭玉.副猪嗜血杆菌的MLST分析及数据库建立[J].南昌大学学报(理科版),2011,35(3):284-287. 被引量:4
  • 10郑念广,谭实勇,何玲,夏芳,高娟,罗满林.副猪嗜血杆菌抗体间接ELISA检测方法的建立与应用[J].中国畜牧兽医,2011,38(8):185-188. 被引量:9

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