关键词: 猪精液, DNA提取方法, 猪细小病毒, 水煮法, 异硫氰酸胍, Trizol方法

Abstract: 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.

Key words: boar semen, DNA extraction, Porcine Parvoviras （PPV）, cooking extraction method, guanidine thiocyanate, Trizol