论文部分内容阅读
建立了牙鲆弹状病毒的环介导等温扩增(loop-mediated isothermal amplification,LAMP)检测方法,首先根据牙鲆弹状病毒的糖蛋白的基因保守序列,利用Primer Explorer V3软件设计了6条引物,并对LAMP反应温度和反应时间等条件进行了优化。该方法的检测限为30fgRNA,比常规RT-PCR灵敏度高100倍,与鲤春血症病毒、传染性胰脏坏死病毒、传染性造血器官坏死病毒、海洋双RNA病毒、病毒性出血败血症病毒以及病毒性神经坏死病毒等没有交叉反应。该方法检测时间短,在1h内即可完成检测。用建立的LAMP方法对临床鱼样进行了检测,结果表明40尾石鲽鱼样品中有3尾感染HRV,与病毒分离结果一致,说明LAMP方法比较适合牙鲆弹状病毒的早期及现场诊断。
A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of Ralstonia solani. Based on the conserved sequence of the glycoprotein of Ralstonia solanacearum, six Primer Explorer V3 softwares were designed. Primers, and LAMP reaction temperature and reaction time and other conditions were optimized. The detection limit of this method is 30fgRNA, which is 100 times higher than that of conventional RT-PCR. The detection limit of the method is 10f, which is similar to that of common carp virus, infectious pancreas necrosis virus, infectious hematopoietic necrosis virus, marine double RNA virus, viral hemorrhagic septicemia virus and Viral nerve necrosis virus did not cross-react. The method of detection time is short, within 1h to complete the test. The established LAMP method was used to detect clinical fish samples. The results showed that there were 3 HRV infected samples in 40 samples, which was consistent with the results of virus isolation, indicating that the LAMP method is suitable for the early and on-site diagnosis of rhabdovirus.