鸡传染性法氏囊病病毒dsRNA的提纯与应用

用传染性法氏囊病病毒（IBDV）攻击4周龄的非免疫鸡，以超速离心法从感染鸡法氏囊组织中分离、纯化IBDV，应用蛋白酶K消化和Trizol（异硫氰酸胍－酚－氯仿法）处理2种方法从纯化的IBDV中抽提dsRNA。通过低熔点琼脂糖割胶－酚－氯仿抽提可获得纯化的dsRNA，研究发现应用蛋白酶K消化获得的纯化RNA产量高，用其作模板进行RT－PCR可扩增IBDV基因组全长cDNA A片段和B片段、VP2基因和VP2－4－3基因。

Extraction of Double-stranded RNA of Infectious Bursal Disease Virus

week old chickens unaffected by infectious bursal disease virus (IBDV) were challenged with Shanghai strains of IBDV orally. The virion were purified through ultracentrifuge from affected bursa. A proteinase K digestion based approach and Trizol RNA extraction method were used to extract dsRNA of IBDV from purified virions. The yield of dsRNA extracted by proteinase K digestion based approach was higher than that of Trizol method. dsRNA extracted by proteinase K digestion was more suitable for the amplification of full length cDNA of IBDV by long and accurate PCR.