| 罗非鱼无乳链球菌巢式PCR检测方法的建立 |
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| 引用本文： | 邵辰,易弋,黎娅,黄荷,伍时华,杨军.罗非鱼无乳链球菌巢式PCR检测方法的建立[J].淡水渔业,2016(2):40-44. |
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| 作者姓名： | 邵辰 易弋 黎娅 黄荷 伍时华 杨军 |
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| 作者单位： | 1. 广西科技大学生物与化学工程学院，广西柳州，545006; 广西糖资源绿色加工重点实验室 广西科技大学，广西柳州，545006; 广西高校糖资源加工重点实验室 广西科技大学，广西柳州，545006;2. 柳州市渔业技术推广站，广西柳州，545006 |
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| 基金项目： | 现代农业产业技术体系专项资金“国家罗非鱼产业技术体系”( CARS －49)；柳州市科学研究与技术开发计划课题(2013E020602) |
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| 摘 要： | 根据无乳链球菌(Streptococcus agalactiae)cfb基因序列,设计、合成2对引物,优化扩增条件,建立了快速高灵敏度鉴别无乳链球菌的巢式PCR方法。结果显示:使用该方法对罗非鱼(Oreochromis mossambicus)血液样品进行检测,可检测到活菌浓度为8.7×104CFU/m L的无乳链球菌。对采自广西地区受无乳链球菌感染的19份罗非鱼样品进行检测,18份可获得目的片段,扩增到的序列均为无乳链球菌cfb基因序列,检测准确度达到94.7%。
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| 关 键 词： | 巢式PCR 无乳链球菌(Streptococcus agalactiae) cfb基因 罗非鱼(Oreochromis mossambicus) |
Development of nested PCR for detection of Streptococcus agalactiae from tilapia |
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| Abstract: | Based on the cfb gene sequence of Streptococcus agalactiae, two pairs of primers were designed and synthesized and the amplification conditions were optimized to establish the nested PCR method which could identify S.agalactiae with high sensitively.The results showed that S.agalactiae could be directly detected from blood sample at a minimum concentra -tion as 8.7 ×10 4 CFU/ml by this method.Nineteen clinical S.agalactiae -infected tilapia samples collected from Guangxi province were applied to verify this method.The results showed that 260 bp fragment could be amplified in 18 samples and the sequencing analysis revealed that all the amplified fragment were cfb gene of S.agalactiae.The detection accuracy of this method was 94.7%. |
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| Keywords: | nested PCR Streptococcus agalactiae cfb gene tilapia (Oreochromis mossambicus) |
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