禽流感油乳佐剂疫苗质量快速检测方法的建立

A Rapid Detection Method of the Quality of Inactivated Oil-adjuvant Emulsion Vaccine against Avian Influenza

通过对低温冻融法、研磨法、稀释高速捣碎法、95%乙醇法及三氯甲烷法破乳效果比较,确定三氯甲烷为油乳剂疫苗的破乳剂。将不同抗原含量的H9和H5亚型灭活与非灭活疫苗破乳后分离上层水相,采用红细胞凝集(HA)试验,测定其HA价,结果发现脱乳后上层水相与未做成油乳疫苗并做相应稀释的原液的HA价下降1～3个滴度,且将不同稀释度制备H5亚型疫苗经脱乳处理后上层水相的HA价与荧光定量PCR结果基本一致。所建立的禽流感油乳剂疫苗质量的检测方法具有快速、经济、简便,尤其适合于对大批量及基层应用。

To demulsify oil-adjuvant emulsion vaccine against avian influenza(AI),repetitive frozen-thaw, grinding at high speed,dilution and mashing at high speed,commingling with of 95% ethanol or chloroform for ratio of 1∶1 were applied,and commingling with chloroform was regarded as demulsifying agent. After various amount of inactivated or live antigen of H5 or H9 subtype AI in oil-adjuvant emulsion vaccine were demulsified,the supernatant was testified by haemagglutination (HA) test for antibody titers. The results showed that the HA titer of supernatant was 1log2 to 3log2 lower than that of corresponding untreated antigen with the same dilution,and the HA titer of H5 subtype AI uaccine was consistent with the results of real-time fluorescence RT-PCR. The method for detection inactivated oil-adjuvant emulsion vaccine against AI was rapid,economical,convenient and cheap,and especially available for detection of large quantities of samples.