CAJ | 학술논문

为研究现用PEDV CV777疫苗毒株对流行毒株的免疫保护效果。以RT‐PCR方法进行PEDV CV777疫苗株、JS12流行株S基因克隆测序，进行PEDV S基因核酸序列分析与氨基酸序列分析。分别以转0瓶工艺与细胞悬浮培养工艺培养Vero E6细胞，制备PEDV CV777株转瓶灭活苗与悬浮灭活苗。分别以PEDV转瓶灭活苗、悬浮灭活苗、PEDV 流行株组织灭活苗（JS12株）产前30 d免疫经产母猪；母猪产仔后14 d进行仔猪攻毒保护试验。核酸序列分析显示：PEDV CV777株S基因与流行毒株的相似性在96．4％～99．7％之间，氨基酸序列相似性在96．7％～99．5％之间。转瓶工艺制备 PEDV 抗原效价为107．0 TCID50／mL ，悬浮培养制备PEDV抗原效价为108．5 TCID50／mL。攻毒保护试验显示：悬浮灭活苗免疫母猪后，仔猪有2／10发病，保护率为8／10，转瓶灭活苗组发病率为8／10，保护率仅为2／10；组织灭活苗组9／10发病；对照组10头全部发病。该试验证明，提高PEDV CV777株的抗原含量，能够有效提高疫苗对PEDV 流行毒株的保护效果。
위연구현용PEDV CV777역묘독주대류행독주적면역보호효과。이RT‐PCR방법진행PEDV CV777역묘주、JS12류행주S기인극륭측서，진행PEDV S기인핵산서렬분석여안기산서렬분석。분별이전0병공예여세포현부배양공예배양Vero E6세포，제비PEDV CV777주전병멸활묘여현부멸활묘。분별이PEDV전병멸활묘、현부멸활묘、PEDV 류행주조직멸활묘（JS12주）산전30 d면역경산모저；모저산자후14 d진행자저공독보호시험。핵산서렬분석현시：PEDV CV777주S기인여류행독주적상사성재96．4％～99．7％지간，안기산서렬상사성재96．7％～99．5％지간。전병공예제비 PEDV 항원효개위107．0 TCID50／mL ，현부배양제비PEDV항원효개위108．5 TCID50／mL。공독보호시험현시：현부멸활묘면역모저후，자저유2／10발병，보호솔위8／10，전병멸활묘조발병솔위8／10，보호솔부위2／10；조직멸활묘조9／10발병；대조조10두전부발병。해시험증명，제고PEDV CV777주적항원함량，능구유효제고역묘대PEDV 류행독주적보호효과。
To carry out the evaluation of the immunogenicity and protective efficacy by inactivated PEDV CV777 vaccines against the epidemic strains .RT‐PCR was used to clone the S gene of PEDV CV777 vac‐cine strain and JS12 strain ,then sequenced the PCR products for analyzing the nuclide sequence and amino acid sequence .PEDV CV777 was prepared by roller bottle technology and microcarrier culture technology in Vero E6 cell separately .Four different groups of sows were used in the attack protection assay :the neg‐ative control group ,the inactivated PEDV CV 777 cultured by roller bottle technology group ,the inactiva‐ted PEDV CV777 cultured by microcarrier culture technology group and the inactivated PEDV JS 12 tissue vaccine group .Sows were vaccinated twice at 30th day before farrowing and then the 14th day piglet was challenged by live virus .Antibody titers in sow and piglet serum sampels were compared by enzyme‐linked immunosorbent assay (ELISA ) . Homology analysis revealed that identity of S gene sequence between PEDV CV777 and the epidemic stains was 96 .4%‐99 .7% ,and the similarity of Amino acid sequence was 96 .7%‐99 .5% .The antigen titer prepared by bottle turning process and microcarrier culture technology were 107 .0 TCID50/mL and 108 .5 TCID50/mL respectively .The attack protection assay proved that the sows immuned by vaccine of microcarrier culture could provide 8/10 protection to piglets ,the vaccines of bottle turning and inactivated tissue provided 2/10 and 1/10 protection ,respectively .Our study indicates that improving the antigen titer will effectively increase the protective efficacy by inactivated PEDV CV 777 vac‐cine against epidemic strains .