| 温和气单胞菌PCR快速诊断方法的建立 |
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| 引用本文： | 黄冠军,刘天强,刘衍鹏,饶朝龙,肖丹.温和气单胞菌PCR快速诊断方法的建立[J].淡水渔业,2012,42(1):89-92. |
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| 作者姓名： | 黄冠军 刘天强 刘衍鹏 饶朝龙 肖丹 |
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| 作者单位： | 通威股份有限公司,成都,610041 |
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| 摘 要： | 根据NCBI公布的温和气单胞菌(Aeromonas sobria)丝氨酸蛋白酶的基因序列,设计并选取一对能够快速而准确地检测温和气单胞菌的PCR引物,建立PCR快速检测体系,并对患病的鱼组织进行检测。研究结果表明,使用设计的引物能够扩增出与预计大小相符合的131 bp的特异性片段,具有较好的检测特异性,对靶标DNA的检测灵敏度为1.0 pg/20μL,对温和气单胞菌的检测灵敏度为50 cfu/20μL。样品的检测结果与实际发病情况一致,表明本研究成功建立了温和气单胞菌常规PCR检测体系,该体系可以用于温和气单胞菌的诊断和样品的检测。
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| 关 键 词： | 温和气单胞菌(Aeromonas sobria) 胞外丝氨酸蛋白酶 聚合酶链式反应 检测 |
Development of a PCR- based rapid diagnosis for Aeromonas sobria |
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| HUANG Guan-jun
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LIU Tian-qiang
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LIU Yan-peng
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RAO Chao-long
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XIAO Dan.Development of a PCR- based rapid diagnosis for Aeromonas sobria[J].Freshwater Fisheries,2012,42(1):89-92. |
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| Authors: | HUANG Guan-jun LIU Tian-qiang LIU Yan-peng RAO Chao-long XIAO Dan |
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| Institution: | (Tongwei Co,LTD,Chengdu 610041,China) |
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| Abstract: | PCR primers were designed to detect Aeromonas sobria rapidly and correctly according to the gene sequence of extracellular serine protease of A.sobria,and one pair of these primers was chosen to establish PCR-based rapid detection system.Samples collected from diseased fish were detected.Results showed that 131bp of expected specific fragment was amplified by the designed primer which had great specificity.The detection sensitivity of the PCR assay was 1.0pg/20 μL for target DNA and 50 cfu/20 μL for A.sobria.The result of the detection was in accordance with the actual situation,which indicated that the PCR-based detection system was developed successfully and can be used to diagnose A.sobria and detect samples. |
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| Keywords: | Aeromonas sobria extracellular serine protease PCR detection |
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