精液中猪细小病毒DNA三种提取方法效果比较

摘要

猪精液中含有高浓度的蛋白质,因此提取高纯度DNA较为困难;试验分别用水煮法、异硫氰酸胍法和Trizol法三种方法对连续稀释梯度猪精液提取猪细小病毒DNA,应用Taqman荧光定量PCR技术评价提取效果并加以对比.结果发现Trizol法可以提取出高纯度的DNA,但因Trizol试剂价格昂贵,不适用临床检测;比较另外两种方法,水煮法较异硫氰酸胍抽提法更为有效,是一种迅速、简便、经济、高效的DNA提取方法,适合于临床大量样品的检测.%To detect Porcine Parvovirus (PPV) in semen,various DNA extraction techniques have been utilized for PCR.but rarely compared,to determine an optimized extraction protocol. Due to the high content of protein,difficulties can be encountered in obtaining DNA from the seminal cell fraction. This study compared three methods of DNA extractions. All extractions were compared on serially diluted seminal cell fractions and evaluated their ability to extract DNA by real-time PCR. The Trizol method resulted in recovery of the highest amount of DNA,but this reagent was quite expensive and was not suitable for routine diagnostic testing. When compared the results of the other two methods,the cooking extraction method was more effective than guanidine thiocyanate method. The analysis result indicated that cooking extraction method was a quick,simple,economic and efficient method for DNA extraction and was suitable for routine diagnostic testing.