【摘要】 将Ⅰ型鸭肝炎鸡胚化弱毒MY株进行纯粹检验;以100ELD50 0.2mL/枚(含病毒3.0×105个拷贝)剂量经鸡胚尿囊腔接种10日龄SPF鸡胚,每隔12h观察并采集鸡胚胚体和尿囊液样本,运用自行建立的检测DHV-1的SYBR GreenⅠ实时荧光定量RT-PCR方法检测MY株弱毒的鸡胚增殖规律.结果表明,该毒种符合我国的兽用生物制品规程,毒种纯粹;病毒接种后,鸡胚死亡时间主要集中在48-53h;病毒在感染后的12h,24h,36h,48h和53h时段,鸡胚尿囊液中病毒拷贝数为:3.28×102、5.28×103、7.01×103、3.31×105和4.41×105;鸡胚胚体病毒拷贝数为:8.46×103、1.91×106、3.41×106、1.36×107和1.03×107;显示,MY株弱毒经鸡胚尿囊腔接种鸡胚,病毒在胚体中的拷贝数在各时间均高于尿囊液中的拷贝数,且在48~53h达到高峰(1.36×107~1.03×107).因此,用鸡胚繁殖MY株弱毒,应以收获鸡胚胚体为主要毒源,收毒时间以鸡胚尿囊腔接种后48~53h为佳.为运用MY株生产I型鸭病毒性肝炎弱毒疫苗奠定了基础.更多还原

【Abstract】 Purity test for duck hepatitis virus(DHV) type Ⅰstrain MY is carried out,and then the strain is inoculated to allantoic cavity of 10 days old chicken eggs at the dosage of 100ELD50 0.2mL per egg containing 3.0×105 copies of the virus.Embryos and allantoic fluid are harvested every 12 hours,and then proliferation profile of strain MY in chicken eggs is investigated with Real-Time RT-PCR for DHV-1 detection which is established by us.The results show that this virus seed is up to the requirements for Veterinary Biological Products in China and the virus seed is pure.The chicken embryo mainly dies at 48 to 53 hours after being inoculated.The copy numbers of the virus in allantoic fluid are 3.28×102,5.28×103,7.01×103,3.31×105 and 4.41×105 at 12h,24h,36h,48h and 53h,while the copy numbers of the virus in embryo are 8.46×103,1.91×106,3.41×106,1.36×107 and 1.03×107.These results indicate that after being inoculated to allantoic cavity of chicken egg,copy number of the virus in embryo is higher than that in allantoci fluid,and reaches the peak at 48 to 53h(1.36×107~1.03×107).Therefore,when strain MY is multiplied with chicken egg,embryo should be harvested as the main virus resource and the best harvest time should be 48 to 53 hours after inoculation.This study provides the foundation for vaccine production by DHV-1 strain MY.更多还原