鸡马立克病活疫苗中污染禽网状内皮组织增生症病毒检测方法的建立

Establishment of indirect immunofluorescence assay method for detection of Avian reticuloendotheliosis virus contamination in Marek's disease live vaccine

为建立鸡马立克病(MD)活疫苗中污染禽网状内皮组织增生症病毒(REV)的间接免疫荧光检测方法,本研究通过向MD活疫苗中添加不同剂量的REV,用拟定的方法对样品检测REV。结果表明,Ⅰ型MD活疫苗、火鸡疱疹病毒活疫苗(Fc-126株)和MD二价活疫苗(HVT+CVI988株)中污染REV的最低检出量分别为每500羽份疫苗中污染5TCID50、10TCID50和15TCID50的REV。应用建立的方法对国内10家企业生产的43批MD活疫苗进行了检验,结果显示,2个企业生产的4批疫苗REV检测阳性。随机选取了5批IFA检测阴性样品和4批IFA检测阳性样品,按鸡检查法进行外源病毒检验,结果两种方法对REV污染检验结果的符合率为100%。因此,本研究建立的IFA法可替代鸡检查法,用于疫苗中REV污染的检验。

To establish an indirect immunofluorescence assay（IFA）method for detection of Avian reticuloendotheliosis virus（REV）contamination in Marek＇s disease（MD）live vaccine,the MD live vaccine was added with different doses of REV and REV was detected by IFA method. The results indicated that the lowest detection limit of I type MD live vaccine（CVI988 or 814 strains）,MD turkeys herpes virus live vaccine（Fc-126 strains）,and MD bivalent living vaccine（HVT＋CVI988 strains）was 5TCID50,10TCID50 and 15TCID50for each500 doses respectively. 43 MD goods vaccines were selected to detect REV contamination using the established IFA method, the results showed 4 batches of vaccine were REV positive,then some positive and negative samples were selected randomly for chicken test, the results indicated that the coincidence rate of the two method was 100%. Accordingly,our research indicated that IFA method could replace chicken test as a safe,easy,and effective method for detection of REV contamination in MD vaccine.