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据对虾白斑综合症病毒(WSSV)的基因保守序列,使用Primer Explorer V3软件设计了2条引物,利用PCR的扩增产物,结合变性高效液相色谱(DHPLC)技术,建立了一种对虾白斑综合症病毒的DHPLC快速检测方法。该检测方法特异性好,与传染性皮下和造血器官坏死病毒、斑节对虾杆状病毒、肝胰腺细小病毒、对虾杆状病毒以及对虾基因组DNA无交叉反应;检测灵敏度较高,约为10-5ng/μL,高于常规PCR及荧光定量PCR的灵敏度。用建立的DHPLC方法对100尾对虾样品进行了临床检测,结果与荧光定量PCR检测结果一致。WSSV的DHPLC检测方法,特异性强、灵敏度高,是对WSSV进行有效检测的一种新方法。
According to the gene conserved sequence of WSSV, two primers were designed by using Primer Explorer V3 software. Based on the amplified product of PCR and DHPLC technology, a white spot syndrome DHPLC rapid detection of disease virus. This assay is specific and does not cross-react with infectious subcutaneous and hematopoietic necrosis viruses, the P. monodon baculovirus, the hepatopancreas parvovirus, the shrimp baculovirus and shrimp genomic DNA; the detection sensitivity is high at about 10- 5ng / μL, higher than the sensitivity of conventional PCR and fluorescence quantitative PCR. The established DHPLC method was used to detect 100 samples of shrimp, and the results were consistent with the results of real-time PCR. WSSV DHPLC detection method, specificity, high sensitivity, is a new method for the effective detection of WSSV.