3种DNA提取方法对WSSV检测结果的影响

he comparison of DNA extraction methods in White Spot Syndrome Virus detection

中文关键词: DNA提取方法  PCR  WSSV

英文关键词: DNA extraction methods  PCR  WSSV

中文摘要:

      比较了酚 氯仿法、煮沸法、试剂盒法3种基因组DNA提取方法对WSSV DNA提取效率、纯度及病毒检测结果的影响。结果表明，3种方法的DNA提取效率平均值分别为 101.5、372.6、21.5 ng/μl；OD260 /OD280范围分别是 1.979~2.175（平均值为2.070）、1.699~1.932（平均值为1.796）、1.784~2.075（平均值为1.951）；OIE巢式PCR阳性率分别为60%、50%、70%；TaqMan定量PCR检测的病毒阳性率均为100%，病毒拷贝含量分别为916.0~2.23×106、63.3~1.78×106、479.7~2.70×106 Copies/μl DNA。

英文摘要:

      In this study, three genomic DNA extraction methods including phenol-chloroform extraction, boiling lysis and reagent kit methods were compared in terms of extraction efficiency, DNA quantity, as well as the results of White Spot Syndrome Virus (WSSV) detection. The DNA extraction efficiency of three methods, was 101.5, 372.6 and 21.5 ng/μl, respectively. The ratios of OD260/OD280 were in the range of 1.979~2.175 (mean 2.070), 1.699~1.932 (mean 1.796) and 1.784~2.075 (mean 1.951), respectively. The positive rate of WSSV by OIE (Office International Des Epizooties) nested-PCR was 60% (DNA from phenol-chloroform extraction method), 50% (DNA from boiling lysis method) and 70% (DNA from kit method), while it was 100% by quantitative PCR (all DNA from three methods). The DNA copy number of WSSV detected by quantitative PCR ranged from 916.0 to 2.23×106, 63.3 to 1.78×106 and 479.7 to 2.70×106 copies/μl, respectively. The results would provide some reference for standardizing the detection of WSSV.