论文部分内容阅读
用地高辛标记正常Ｖｅｒｏ细胞ＤＮＡ，制成特异探针，通过与制备人用狂犬疫苗的基质Ｖｅｒｏ细胞同源ＤＮＡ杂交，检测人用狂犬疫苗中残余Ｖｅｒｏ细胞ＤＮＡ含量，达到ＷＨＯ规程中“以传代细胞制备生物制品中残余细胞ＤＮＡ含量，每剂量ＤＮＡ须低于１００ｐｇ”的检测要求。检测灵敏度可达１ｐｇ，杂交反应特异性高，重复性好。测得以Ｖｅｒｏ细胞制备人用狂犬疫苗粗制原液中残余细胞ＤＮＡ含量为１００～１０００ｐｇ／ｍｌ，超滤浓缩后为１０４～１０６ｐｇ／ｍｌ，纯化后低于１０～１００ｐｇ／ｍｌ。这对以传代细胞为基质的生物制品中残余细胞ＤＮＡ的检测提供了一种常规可行的方法，因而具有重要的应用价值
Digoxigenin was used to label DNA of normal Vero cells to make specific probe. The DNA content of residual Vero cells in rabies vaccine for human was detected by homologous DNA hybridization with matrix Vero cell prepared from human rabies vaccine, Cell preparation of biological products in the residual cell DNA content, each dose of DNA should be less than 100pg "detection requirements. Detection sensitivity up to 1pg, hybridization reaction specificity, good repeatability. The residual DNA content of crude rabies vaccine prepared from Vero cells was 100-1000 pg / ml, followed by 104-106 pg / ml after ultrafiltration and less than 10-100 pg / ml after purification. This provides a routine and viable method for the detection of residual cellular DNA in cell-based biological products and therefore has important applications