论文部分内容阅读
选择派琴虫保守的核糖体DNAITS-2区域设计引物和TaqMan探针,通过对反应体系和反应条件进行优化,建立了实时荧光定量PCR检测派琴虫的方法。所构建方法检测质粒模板DNA的动态范围为2.6×101～2.6×107拷贝,敏感度可检测到26拷贝质粒DNA,而且与包拉米原虫、隐孢子虫等其他寄生性原虫无交叉反应,也不受贝类组织DNA的干扰。利用本研究所建立的方法对来自我国山东、福建等不同沿海海域的30份贝类样品进行检测,检出阳性样品3份。研究表明,本研究所构建的派琴虫实时荧光定量PCR检测方法具有快速、灵敏和特异等优点,可满足国内养殖场及进出口水生动物携带派琴虫的检测需要。
The method of real-time fluorescence quantitative polymerase chain reaction (PCR) was established for the detection of fusiformis by optimizing the reaction system and reaction conditions by selecting the conserved ribosomal DNAITS-2 region primer and TaqMan probe. The constructed plasmid DNA showed a dynamic range of 2.6 × 101 ~ 2.6 × 107 copies, 26 copies of plasmid DNA could be detected with sensitivity and no cross-reaction with other parasitic protozoa such as Bombyx malaria and Cryptosporidium, Not affected by shellfish DNA interference. Using the method established in this study, 30 samples of shellfish from different coastal areas in Shandong, Fujian and other coastal areas were detected and 3 samples were positive. The research shows that the real-time fluorescence quantitative PCR detection method of fidiamania collected in this study is fast, sensitive and specific, which can meet the detection needs of domestic fishes and import and export aquatic animals.