为探究羊经布鲁菌病疫苗(S2株)免疫后的血清抗体长期动态消长规律,本试验于2012年9月-2015年9月选取阿拉善盟阿拉善左旗96只山羊、100只绵羊作为试验动物,经布鲁菌疫苗(S2株)100亿CFU、200亿CFU不同免疫剂量灌服,分别于免疫前和免疫后不同时间段采血分离血清,应用虎红平板凝集试验和试管凝集试验进行抗体监测.结果 表明,绵羊组在一免后20 d时,血清抗体阳性率达到最高峰,之后呈下降趋势,150 d抗体阳性率降至0％,免疫后180 d ～360 d,绵羊组抗体阳性率一直处于较低水平(0％～4％),二免和三免后不同时间段免疫抗体消长规律与首次免疫基本一致;山羊组抗体阳性率在免疫后20 d时,血清抗体阳性率达到最高峰,之后呈下降趋势,免疫后90 d～360 d,免疫抗体时高时低,起伏较大,无明显规律,二免和三免后不同时间段免疫抗体消长规律与首次免疫基本一致,结果同时表明,100亿CFU和200亿CFU两个免疫剂量组的抗体阳性率无明显差异.

In order to evaluate the effect of control measures on brucellosis in Inner Mongolia and to further polish con-trol strategy, an evaluation was conducted on different control measures that being implemented in Siziwangqi county. The targeted control measures included tracing and culling strategy, vaccination, surveillance, quarantine and so on. The result showed that the tracing and culling strategy has distinctive effect in general, but the effect might vary along with different infection statuses. Single tracing and culling was unable to remove all the infected individuals when the prevalence was high, while“vaccination without tracing and culling strategy” had a certain effect on brucellosis control in that situation.

To investigate the pattern of increasing or decreasing of serum antibody in sheep and goats immunized with brucellosis vaccine,RPST and tube agglutination test were used in the study to determine the level of serum antibody in sheep and goats before and after immunization with Brucella strain S2 vaccine. The results indicated that the serum antibody positive rate in sheep reached the highest level 20 DPI,then started to decrease and remained relatively low level 90-360 DPI,mostly below 6%,and relatively higher in few individuals,up to 14%. The positive rate in goats reached the highest 20 DPI,then decreased and remained low or relatively low level 90-360 DPI,but with decreasing or increasing sometimes. Comparatively,antibody positive rate in goats was generally higher than in sheep,and the descending and ascending range was bigger. It was also shown that there was no significant difference in antibody posi-tive rate between 10 billion CFU dose group and 20 billion CFU dose group in same species.

为准确掌握牛羊口蹄疫疫苗免疫效果,2012年对内蒙古自治区的牛羊口蹄疫O-Asia I型双价灭活疫苗和牛羊口蹄疫A型灭活疫苗免疫效果进行了监测.采用液相阻断ELISA方法,共检测了8个旗县23018份免疫牛羊口蹄疫O-Asia I型双价灭活疫苗和牛羊口蹄疫A型灭活疫苗15日、30日、60日的血清中口蹄疫免疫抗体水平.结果显示,牛羊口蹄疫疫苗免疫效果总体良好,免疫抗体合格率超过了70％.规模化奶牛养殖场免疫效果稳定,散养地区免疫效果稳定性差,且不同厂家的疫苗免疫效果不同,加强免疫后A、B、D三个厂家抗体合格率能达到100％,C厂家的免疫效果较差,抗体合格率为93.3％.采用3ABC-ELISA检测试剂盒对部分地区牛羊进行感染抗体检测,结果为阴性.

To analysis the sensibility, specificity and coincidence rate of the complement fixation enzyme linked immunosorbent assay kit ( CF-ELISA kit ) for detecting brucellosis antibodies, 400 bovine and sheep serum specimens were collected from brucellosis-affected areas selecting CF-ELISA kit to detect brucellosis antibodies, where cases had already been identified. Another total of 200 bovine and sheep serum specimens for detecting antibodies were collected from no brucellosis-affected areas, where no cases had been identified. Results showed that there was no significant difference (P>0.05) among the CF-ELISA kit, iELISA kit and CGS for detecting antibodies of brucellosis-affected areas serum. Kappa statistics result showed that the method of CF-ELISA and iELISA had highly coincidence rate. The specificity and coincidence rate were 100% between the CF-ELISA kit and iELISA kit for detecting antibodies of no brucellosis-affected areas serum.

【Objective】 The study was conducted to express nonstructural protein VPg1-2,VPg2-3,VPg3 of the foot-and-mouth disease virus(FMDV) for differentiation of the infected and vaccinated animal【Method】VPg1-2(3B1-2),VPg2-3(3B2-3),VPg3(3B3) gene fragments were successfully amplified by PCR and subcloned from 3ABC gene of the foot-and-mouth disease virus(FMDV) Taibao strain.Their expressing plasmids were constructed by inserting the target gene fragments into pGEX-4T-1 vector.The expressed proteins were analyzed by Western blotting.An indirect ELISA was set up using purified VPg1-2 protein as antigen,and 4 groups of background-known cattle sera were tested.【Results】The expressed VPg1-2,VPg2-3 proteins reacted with FMDV positive serum while the reaction of the VPg3 protein was negative.Negative results were obtained for the non-vaccinated control group and the vaccinated group and a part of the infected post-vaccination group(3/7).On the contrary,positive results were achieved for the infected group and a part of the infected post-vaccination group(4/7).In the infected group and infected post-vaccination group,the longest duration of the antibody against VPg1-2 lasted for at least 90 days.The earliest appearing time of VPg1-2 antibody was10 days after infection.【Conclusion】The expressed VPg1-2 proteins’sspecificity and sensitivity were 100% for differentiating infected from vaccinated in the non-vaccinated and the vaccinated and the infected cattle.May be it’s lower level of sensitivity for the infected post-vaccination cattle.

The VP0,VP3,VP1 genes of Avian encephalomyelitis virus Van Roekel strain were successfully cloned and VP0,VP3,VP1 expressing plasmids were consturcted by inserting the target genes into pGEX-4T-1 vectors respectively.The VP1 expression proteins were analysed by ELISA. The result showed that expressed VP1 protein reacted with AE positive serum more stronger than the other two. An indirect ELISA method was set up using purified VP1 protein. as antigen and tested 10 serum samples. The result shown that 8 samples of them were positive,their S/P level are higher than 0.2.At the same time,10 serum samples were detected by ELISA kit and AGP.The results of detection showed that 8 of them were positive,but others are negative.This proved that the activity of expression protein satisfied the anticipative need.The result showed, this method can be used for diagnosis of AE.

VPg2 gene fragment was successfully subcloned from 3ABC gene of the foot-and-mouth disease virus(FMDV) TaiBao strain and constructed VPg2 expressing plasmid by inserting the target gene fragment into pGEX-4T-1 vector.The expressed protein was analysed by Western blotting.The result showed strong reaction with FMDV positive serum.VPg2 protein ELISA was evaluated through testing sera of 4 cattle groups i.e.control group,vaccinated group,infected group and infected after vaccination group.Negative results were obtained for the control group and the vaccinated group.On the contrary,positive results were obtained for the infected group and part of the infected after vaccination group.In the infected group and infected after vaccination group,the duration of the antibody against VPg2 lasted for at least 90 days.The earliest detecting time of VPg2 antibody was 10th day after infection.The duration of the antibody against VPg2 was the same with the antibody against 3ABC tested previously.2170 negative sera were tested using indirect VPg2-ELISA and 3ABC-ELISA.The results showed that the false positive rates were 0.55% in VPg2 ELISA assay and were 2.21% in 3ABC-ELISA assay.

成功克隆到O型口蹄疫病毒（FMDV）太保毒株3ABC基因片段中的3A、3B、3C基因,并将它们插入pGEX-4T-1或24B11表达载体构建了重组表达质粒,经Westernblotting分析表明,表达的3A、3B蛋白可与FMDV阳性血清发生特异性反应,但表达的3C蛋白没有反应。以纯化的3A、3B表达蛋白为抗原建立间接ELISA,对4组背景清楚的试验牛血清进行检测,结果表明3A、3B表达蛋白与非免疫对照组（C组）和灭活疫苗反复免疫组（CI组）牛血清均不发生反应,与未免疫直接攻毒组（D组）和免疫后攻毒组（I组）牛血清均发生反应;D组和I组3A、3B表达蛋白抗体持续时间最长可达90d以上,3A和3B表达蛋白最早检出相应抗体的时间分别为攻毒后第5天和第10天。

The experimental avian encephalomyelitis virus (AEV) strain was obtained by means of AEV being passaged in the SPF avian embryos. According to the published sequences of VP0, VP3 and VP1 genes of AEV strain 1143, 3 pairs of primers were designed. The cloning and sequence analysis of the VP0, VP3 and VP1 genes of AEV VR strain was carried out after the VP0, VP3 and VP1 genes of AEV VR strain were amplified respectively with the 3 pairs of primers by RT-PCR. The results showed that the VP0, VP3 and VP1 genes of AEV VR strain shared respectively 95.1%,93.6% and 93.9% homology with those from AEV strain 1143, and that the deduced amino acids sequences from the VP0,VP3 and VP1 genes of AEV VR strain shared respectively 97.5%,97.5% and 99.6% homology with those from AEV strain 1143.

A gene fragment of 3ABC gene of the foot-and-mouth disease virus (FMDV) TaiBao strain was successfullycloned, and 3ABC, 3AB, 3BC expressing plasmids were constructed by inserting the target gene fragments into pGEX-4T-1 vectors respectively. The expression proteins were analysed by Western blot methods. The result showed that theexpressed 3ABC and 3AB proteins strongly reacted with FMDV positive serum while the reaction of the 3BC protein withthe FMDV positive serum was poor. An indirect ELISA was set up using purified 3ABC and 3AB proteins as antigen andtested against 4 groups of cattle serum: control group, vaccinated-group, infected-group and infected-group after vaccination.Negative results were obtained using the control group, the vaccinated group and a part of the infected-group aftervaccination. On the contrary, positive results were obtained in the infected group and a part of the infected-group aftervaccination. In the infected group, the duration of the antibody against 3ABC and 3AB were the same and have lasted forat least 90 days. The duration of the antibody against 3ABC is longer than that against 3AB in the infected-group aftervaccination. This indicated that the expression 3ABC protein is more suitable for using as an antigen to differentiate theconvalescent and vaccinated animals.

在内蒙古地区5个种鸡场采血、分离血清共229份,其中3个肉种鸡场血清共184份;2个蛋种鸡场血清共45份,在ALV-J gp85抗原包被的96孔板中进行微量滴定,检查结果,ALV-J抗体阳性率:3个肉种鸡场分别为28.12%,9.38%和8.33%,平均为12.0%;2个蛋种鸡场都为0%.

We investigated the infection of \%Avian leukosis virus\% subgroup J myelocytomatosis using ELISA method in Inner Mongolia Region. The result indicated that the meat type flocks of three different farms infection rate of ALV J were 28.12%(9/32)?9.38%(3/32)and 8.33%(10/12),respectively,and all laying chickens infection rate were 0%(0/35,0/10).We chose 6 chickens of ALV J antibody positive and 13 chickens of ALV J antibody negative to do PCR test. The result indicated that 6 chickens of ALV J antibody positive were all PCR positive,and 4 chickens were PCR positive among 13 chickens of ALV J antibody negative. The result indicated that there were ALV J virus not only in ALV J antibody positive chickens but also in some ALV J antibody negative chickens.