| 副猪嗜血杆菌实时荧光PCR快速检测方法的建立和应用 |
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| 引用本文： | 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):3607-3609,3629. |
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| 作者姓名： | 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 |
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| 作者单位： | 广西兽医研究所,广西南宁,530001 |
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| 摘 要： | 目的]建立一种快速准确定量检测副猪嗜血杆菌（HPS）的检测方法。方法]根据GenBank公布的副猪嗜血杆菌16 S rRNA基因序列,选择其保守区域设计1对特异性引物。通过优化引物浓度和退火温度,建立快速定量检测副猪嗜血杆菌的实时荧光PCR方法,并评价了其特异性和稳定性。结果]建立的实时荧光PCR方法最低检测限是50拷贝/μl,重复性好,变异系数均小于2%。该方法特异性强,只能检测副猪嗜血杆菌,不能检测到猪链球菌2型、金黄色葡萄球菌、大肠杆菌DH5α和猪伤寒沙门氏菌。采用该方法对10份临床疑似HPS感染样本进行检测,其结果与细菌分离培养和常规PCR的结果一致。结论]建立的实时荧光PCR方法快速、灵敏、特异、重复性高,可用于HPS感染的临床快速检测。
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| 关 键 词： | 副猪嗜血杆菌 实时荧光PCR 16S rRNA |
Development of Real-Time Fluorescent PCR for Rapid Detection of Haempohlius parasuis |
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| LI Jun et al.Development of Real-Time Fluorescent PCR for Rapid Detection of Haempohlius parasuis[J].Journal of Anhui Agricultural Sciences,2011,39(6):3607-3609,3629. |
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| Authors: | LI Jun |
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| Institution: | LI Jun et al(Guangxi Veterinary Research Institute,Nanning,Guangxi 530001) |
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| Abstract: | Objective] To develop a real-time fluorescent PCR assay for rapid detection of Haempohlius parasuis(HPS).Method] According to the conservative sequences of 16 S rRNA genes of HPS published in GenBank,a pair of specific primers was designed.The real-time fluorescent PCR was developed by optimizing primer concentration and annealing temperature.And its specificity and reproducibility were evaluated.Ten HPS-suspected samples were detected by the developed method.Result] The lowest detection limit of the dev... |
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| Keywords: | 16 S rRNA |
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