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聚合酶链式反应检测猪细小病毒方法的研究
Establishment of polymerase chain reaction(PCR) for detecting porcine parvovirus(PPV)
【摘要】 本研究依据编码猪细小病毒结构蛋白VP2基因序列,合成了一对寡核苷酸引物,通过减少病毒核酸的提取时间、费用及优化聚合酶链式反应(PCR)的条件,成功地从猪细小病毒(PPV)感染的细胞中扩增出预期的158bp片段,经EcoRⅠ酶切鉴定,证实了该扩增片段的特异性。经敏感性试验测定,PCR的最低检出量为0 008病毒血凝(HA)单位。这些结果表明本试验建立的PCR对PPV的检测人有快速、简便、经济、灵敏度高和特异性强的特点。
【Abstract】 A pair of 20base primers within PPV structure vp2 coding region was synthesized.Through the decrease of time and cost of extracting DNA from PPVinfected PK15 cells and optimizing PCR conditions,the expected 158?bp fragment was amplified.The amplified fragment was shown to be specific for PPV DNA as indicated by EcoR Ⅰ digestion.These results showed the developed PCR is a fast、simple、economical、specific and sentitive detection method.