也谈生命 联系站长

浏览量

鼠疫耶尔森氏菌三色荧光定量PCR检测要领的成立

作者:互联网 发布时间:2016-12-12

    潘光合1,谭勇1,吴文旺1,甘洁1,梁亮2,覃杨光1,王鸣柳3,万道正2,蓝翊文2,梁中平2

  1.广西进出境检讨检疫局,广西 南宁 530021;2.凭祥进出境检讨检疫局;

  3.广西壮族自治区疾病防范节制中心

  摘要:目标  成立一种三色荧光定量PCR检测鼠疫耶尔森氏菌的要领。要领  按照鼠疫耶尔森氏菌pPCP1质粒上的pla基因、编码F1抗原在pMT1质粒上的ca f1基因以及菌体染色体上的3a基因序列,设计特异性的引物和差异荧光标志的Taqman荧光探针,六氟锑酸钠,优化回响体系和扩增条件,耐磨材料,考查该要领检测的敏感度、特异性、反复性和不变性,最终成立三色荧光定量PCR检测要领。功效  本研究所成立的要领对鼠疫耶尔森氏菌DNA的扩增效率高,最低检出限为1×102 copies/ml,特异性高,硼化铁,反复性的变异系数在公道范畴内,不变性好。结论  成立了一种同时检测鼠疫耶尔森氏菌pla、ca f1和3a基因的三色荧光定量PCR要领,该要领敏感度和特异性高,可以或许实现快速检测的目标。

  要害词:三色荧光;定量PCR要领;鼠疫耶尔森氏菌;检测

  中图分类号:R446  文献标识码:B

  Establishment of a three-color fluorescence quantitative PCR method

  for detecting Yersinia pestis

  PAN Guang-he﹡, TAN Yong, WU Wen-wang, GAN Jie, LIANG Liang, QIN Yang-guang, WANG Ming-liu,

  WAN Dao-zheng, LAN Yi-wen, LIANG Zhong-ping

  ﹡Guangxi Entry-exit Inspection and Quarantine Bureau, Nanning, Guangxi 530021, China

  Abstract: Objective   To establish a three-color fluorescence quantitative PCR method to detect Yersinia pestis.   Methods   The specific primers and Taqman fluorescent probes were designed according to pla gene on pPCP1 Plasmid, ca f1 gene on pMT1 Plasmid and 3a gene on chromosome of Yersinia pestis. The reaction system and amplification conditions were optimized. The sensitivity,碳化钛铝, specificity,repeatability and stability of the method were studied. The three-color fluorescent quantitative PCR method for detecting Yersinia pestis was eventually establishred. Results  The detecting ratios and amplification efficiency of the method for samples of Yersinia pestis DNA was high, the line detecting limit of the method was 1×102 copies/ml, with better specificity,硬脂酸镍, the repeatability was in the reasonable range, and the stability was good. Conclusion  The three-color fluorescent quantitative PCR method to detect the pla, ca f1 and 3a gene of Yersinia pestis is highly sensitive and specific, and can be used to detect Yersinia pestis rapidly.

  Key words: Three-color fluorescence; Quantitative PCR; Yersinia pestis; Detection

  《中国国境卫生检疫杂志》2014年3期

  

网友吐槽

推荐阅读

创业故事

友情链接

前沿科技 高新技术 一路同行 技术动态 也谈生命

纳米材料,纳米技术,纳米科技,纳米论坛,中国纳米技术,新材料