实时荧光定量PCR及RT-PCR与常规PCR及RT-PCR检测对虾病毒效果的比较

常规PCR及RT-PCR已用于对虾DNA及RNA病毒检测,但存在费时、灵敏度较低、不能定量等问题。建立了TaqMan实时荧光定量PCR及RT-PCR方法,分别用于检测白斑综合症病毒(WSSV)、传染性皮下及造血组织坏死病毒(IHHNV)及桃拉综合征病毒(TSV)、黄头病毒(YHV)4种对虾病毒。与常规PCR及RT-PCR比较,所建立的TaqMan实时荧光定量PCR及RT-PCR检测上述4种对虾病毒不仅有很高的特异性,检测灵敏度也提高了10～100倍,同时还具有快速、简便、不污染环境、重复性好、实时定量等优点,可明显提高对虾病毒检验检疫工作质量及效率。

Comparison of the Effects among Real-Time Fluorescent Quantitative PCR and RT-PCR with Routine PCR or RT-PCR to Detect Prawn Viruses

Routine PCRs or RT-PCRs have been used to detect prawn DNA and/or RNA viruses, however, they have some problems such as time - consuming, poor sensitivity and unable to detect by quantity etc. In the present study, Taqman real-time fluorescent quantitative PCRs or RT-PCRs were established to respectively detect white spot syn- drome virus （WSSV） and infectious hypodermal and hematopoietie necrosis virus （IHHNV） or Taura syndrome virus （TSV） and yellow-head virus （YHV） in prawn. As compared with routine PCR or RT-PCR, the established TaqMan real-time fluorescent quantitative PCRs Or RT-PCRs to test the above-mentioned four diseases not only had higher de- tection specificity, but also had higher sensitivity by 10 to lO0-fold increment of detection, at the same time, it had the advantages such as rapid, convenient, non-environment-polluted, good repeatability, real-time and quantitative, which could remarkably improve the quality and efficiency of the inspection and quarantine against the prawn viruses.