兔抗猪肺炎支原体胞膜和胞浆抗原的抗体效价测定

将猪肺炎支原体胞膜和胞浆抗原接种家兔后10－14天，可检出抗体，4周后血清抗体效价都达到一定高度，并维持到21周，以后逐渐下降，到23周时仍能检出抗体。     

口蹄疫疫苗非抗原蛋白对146S抗原免疫效果的影响

为探究非抗原蛋白对口蹄疫病毒A型(FMDV-A) 146S抗原免疫效果的影响,本研究以小鼠和猪为试验动物,分别制备5组小鼠注射用样品:A组(3μg 146S抗原)、B组(3μg 146S抗原+25μg非抗原蛋白)、C组(3μg 146S抗原+50μg非抗原蛋白)、D组(3μg 146S抗原+100μg非抗原蛋白)及E组(空白对照,PBS);同时制备4组猪注射用样品:A组(18μg 146S抗原)、B组(18μg 146S抗原+400μg非抗原蛋白)、C(18μg 146S抗原+4 000μg非抗原蛋白)及D组(空白对照,PBS)。免疫接种试验动物后,通过淋巴细胞增殖试验及实时荧光定量PCR评价比较不同疫苗样品组小鼠产生的细胞免疫应答水平,同时通过液相阻断ELISA方法检测、评价各试验组动物(小鼠和猪)产生的体液免疫应答水平。小鼠试验结果显示,在免疫后3个检测时间点内,4个试验组中C组的平均抗体水平最高,平均抗体效价分别为4.13、5.83和5.50,而D组的抗体水平最低,平均抗体效价分别为3.46、5.16和4.46;C组细胞增殖能力及Th-1型细胞因子IL-6、IFN-β和TNF-αmRNA表达量均高于其他组。猪试验结果显示,A组和B组间平均抗体水平差异不显著(P>0.05),但A组和B组的平均抗体效价均极显著高于C组(P<0.01)。综合上述结果表明,一定范围内少量非抗原蛋白对FMDV-A 146S抗原的免疫效果没有影响,而高浓度的非抗原蛋白则抑制FMDV-A 146S抗原的免疫效果。     

旋毛虫S_3抗原的提取及其抗独特型抗体制备

通过匀浆、差速离心等方法提取旋毛虫肌幼虫S3抗原,并对其进行了琼脂双向双扩散、免疫电泳、聚丙烯酰胺梯度凝胶电泳、薄层等电聚焦电泳、肌幼虫孵化试验等免疫学和生化特性分析。以S3抗原免疫兔,再将其抗体免疫同种异体兔,经亲和层析共获得3株抗独特型抗体(R1、R2、R3)。固相放射免疫竞争抑制试验表明,随着抗独特型抗体浓度的升高,其对125I标记的S3抗原-兔Ab1系统及人Ab1系统抑制率均升高,其中R1株对以上两系统的最高抑制率分别达92%和68%。进而证实,3株抗独特型抗体可变区结构与旋毛虫肌幼虫S抗原的决定簇或其邻近结构是相似的,为种间交叉反应型抗独特型抗体。     

泰万菌素对母猪群蓝耳病的治疗试验

本文以泰万菌素为对象,研究其对母猪蓝耳病的治疗效果。实验以猪蓝耳病阳性不稳定场母猪群600头,对比用药前后母猪抗原抗体水平及各项生产性能指标变化。结果显示：用药前后PRRSV抗原阳性率由86.7%降到8.3%;猪群PRRSV抗体水平明显改善,S/P值在0.4～2.0范围由43.3%提升到95.0%,S/P值整体水平差异显著（p<0.05);母猪群各生产性能均得到改善,恢复到正常水平。说明泰万菌素对母猪蓝耳病治疗效果明显,提升猪群健康水平。     

囊虫病猪血清CA效价与虫负荷的相关性及CA与Ab的动态观察

应用ELISA双抗体夹心法检测囊虫病猪血清中的循环抗原(CA),对CA效价与虫负荷的相关性作了初步观察,发现两者密切相关.轻度感染猪血清CA效价≤000,中度为1000～10000,重度为10000～100000.人工感染病猪血清一般在感染后1周检出CA,于感染后4～5周达高峰;抗体一般于感染后2周检出.这种方法可检出的最小虫负荷约为20个/猪     

猪传染性胃肠炎病毒S基因B和C抗原位点片段克隆序列分析及原核表达

猪传染性胃肠炎病毒(TGEV)S蛋白可诱导中和抗体，是主要保护性抗原，N端抗原位点B和C在猪呼吸道冠状病毒(PRCV)中缺失。体外扩增TGEVS基因B、C抗原位点357bp片段(TS)。核苷酸序列与氨基酸同源性分析表明该片段较为保守。将TS克隆到原核表达载体pGEX-6P-1中，转化大肠杆菌中。经IPTG诱导后，SDS—PAGE分析目的蛋白与谷胱苷肽硫转移酶(GST，大小约为26ku)融合后大小约为40ku，目的蛋白分子量约为13．2ku，优化表达条件后表达量达37．9％。     

应用猪囊虫匀浆抗原间接血凝试验检测猪囊虫病抗体的研究

从囊虫匀浆浸提液中分离出一种抗原性与特异循环抗原相同的成分,以这种抗原成分通过间接血凝试验检测病、健猪的血清抗体,结果病猪抗体阳性率为93.2％(96/103),健猪阴性(?)合率为100％。     

旋毛虫感染小鼠对p46 000重组抗原的抗体应答

分别以旋毛虫肌幼虫ES抗原和p46000重组蛋白作为抗原，对小鼠人工感染旋毛虫后的抗体应答进行了ELISA检测。结果表明，以肌幼虫200条／只经口感染小鼠后，肌幼虫ES抗原在感染后9d可检出抗体，并于感染后35～42d达到最高水平；应用重组抗原检测时，感染后10d可检出抗体，抗体水平略低于用ES抗原，但是其消长规律基本一致．而且与阴性血清相比差异明显；抗体在117d后仍维持于较高水平。     

猪流行性腹泻病毒S1基因的截段克隆表达及其在卵黄抗体制备上的应用

通过RT-PCR技术扩增了猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)S1截段基因序列,将扩增的S1基因截段克隆到原核表达载体pET-32α(+)中,构建了重组表达质粒pET-32α-S1,经测序鉴定正确后,转化至感受态细胞BL21(DE3)中,并进行IPTG诱导表达。结果显示:重组菌可表达相对分子质量为45 000的融合蛋白,蛋白表达量较高;Western blotting结果显示,表达的S1蛋白能与PEDV阳性血清发生特异性的免疫反应,表明原核表达的S1蛋白具有良好的反应原性;将纯化后的S1蛋白与蜂胶佐剂按比例混合,作为免疫抗原,免疫产蛋鸡,每隔2周免疫1次,共免疫3次,定期利用ELISA方法检测抗体效价水平,琼脂扩散试验抗体效价最高达到1∶64,收集并纯化卵黄抗体(IgY),SDS-PAGE结果分析纯化效果较好;在人工感染仔猪试验中,卵黄抗体试验组的治愈率为100%,说明卵黄抗体对猪流行性腹泻病具有较好的治疗效果。结果表明:表达的重组S1蛋白具有良好的抗原性,作为免疫抗原免疫产生的卵黄抗体具有较高的抗体效价,为进一步研制猪流性腹泻卵黄抗体生物制剂奠定了基础。     

犬瘟热病琼脂扩散诊断方法的建立

本文主要阐述了用鸡胚成纤维细胞培养的犬瘟热弱毒疫苗免疫绵羊制备琼扩抗体，用犬瘟热病貉肝脏制备琼扩抗原，建立了琼脂扩散诊断方法。用于犬瘟热死亡动物的肝、脾脏器病毒抗原检出，并初步检测送检病科１０份，与电镜检查结果比较，阳性符合率８５．７％，阴性符合率７５％，总体符合率为９０％。方法特异性强，敏感性高，操作易行，便于推广应用。     

两种免疫酶试剂盒检测猪抗E型肝炎病毒IgG的比较

目的比较两种免疫酶试剂盒recomWell HEV IgG(swine)ELISA(recom Well )和recomLine HEV IgG(swine)(recom Line )检测猪抗E型肝炎病毒(hepatitis E virus,HEV)IgG的准确性。方法分别采用基因3型和基因4型人HEV(human HEV,hHEV)人工感染无菌猪各2头,采用上述试剂盒检测猪感染后不同天数的血清样品中抗HEV IgG,同时对两头接种PBS液的无菌猪血清样品18份和4头未感染猪的18份血清进行了检测。结果 2头猪人工感染基因3型hHEV后,recomWell检测均于感染后21d出现阳性,并持续56d;recomLine检测,其中1头猪于感染后14d呈阳性,而另1头于感染后21d呈阳性,且均持续56d。2头猪人工感染基因4型hHEV后,两种试剂盒检测,其中1头猪于感染后21d呈阳性,另1头35d呈阳性,并均持续56d仍为阳性。18份对照血清两种试剂盒检测均为阴性。18份未人工感染猪血清中,只有1份两种试剂盒检测均为阳性。两种试剂盒同时检测的共72份样品中,recomWell检出阳性样品23份,阳性率约为32.0%(23/72),recomLine检出阳性样品24份,阳性率约为33.30%(24/72),检出阳性率两者差异不显著(P>0.05);在recomWell检出的23份阳性样品中,recomLine检测均为阳性,两者阳性检出符合率为100%(23/23);recomWell的漏检率约为4.1%(1/24)。结论 recomWellR和recomLine均可用于猪抗HEVIgG的检测,recomLine检测灵敏度略高于recomWell,且操作更简便,不需要特殊昂贵的检测设备,特别适合用于基层检测猪抗HEV IgG。     

贵州奶牛传染病检测和预防的研究

2006—2008年对贵州省内610份奶牛血清进行检测,其中有116份血清为乙型肝炎表面抗原(HBsAg)阳性,平均阳性率19%;534份牛血清样品中,检出奶牛传染性鼻气管炎阳性血清29份,阳性率为5.4%,而运用血清补体结合反应(CFT)检测奶牛传染性胸膜肺炎,均为阴性。将HBsAg、HBeAg双阳性奶牛肝组织做电镜观察,发现肝组织中有密集的HBV颗粒。结果表明,贵州省奶牛可能存在乙型肝炎、传染性鼻气管炎的感染,应当引起奶牛养殖者的高度重视。     

A SEROLOGICAL SURVEY FOR BLUETONGUE VIRUS ANTIBODY IN WESTERN AUSTRALIA

A serological survey was carried out to detect specific (serotype 20) and a group bluetongue virus antibody in cattle and sheep serums collected in Western Australia during the period January 1 1978 to June 30 1979. Of 18,849 cattle serums examined by the gel diffusion precipitin test (GDPT), 9.7% were positive and 6.1% gave doubtful results. All 1949 sheep serums tested were negative. Precipitin antibody was demonstrated in 22.5% of serums from Kimberley cattle and 3.6% of cattle serums from the Northwest. Serums collected from cattle in the South were consistently negative in GDPT. When 915 serums that reacted in the GDPT were further tested by the complement fixation test (CFT), 164 were positive. The percentage of CFT positive serums increased as the GDPT reaction became stronger. 2467 serums collected from cattle in Kimberley and Northwest areas and tested by the CFT, 175 (7.1%) were positive. These 175 positive serums were also examined by GDPT and 164 doubtful or positive reactions were obtained. The virus neutralisation (VNT) using serotype 20 virus was carried out on 3804 serums, including all serums that reacted in the GDPT, and 57 were positive. When the VNT positive serums were examined in the other 2 tests, 47 serums were either positive or doubtful in the GDPT and 8 were positive in the CFT. The presence of bluetongue virus group antibody in cattle serums closely followed the suggested distribution pattern of Culicoides brevitarsis but specific serotype 20 neutralising antibody was limited to cattle serums from stations situated north of latitude 17 degrees S in an area of mean annual rainfall higher than 700 mm.     

A SURVEY OF ANTIBODY TO AINO VIRUS IN CATTLE AND OTHER SPECIES IN AUSTRALIA

SUMMARY A serological survey of healthy cattle in Australia showed that antibodies to Aino virus were present in serums from cattle in northern Australia and down the east coast as far as central New South Wales in 1975, 1976 and 1977, but occurred with a lower frequency than antibodies to Akabane virus. in contrast to the findings with Akabane virus, no neutralising antibodies to Aino virus were detected in serums from camels, dogs or horses. Antibodies to both viruses were detected in buffaloes and sheep, but not in humans or any of the Australian indigenous species so far tested. All positive serums originated from within the known range of Culicoides brevitarsis.     

Applied serology in the latter stages of the eradication of bovine brucellosis

Late in the program to eradicate bovine brucellosis from Western Australia, Rose Bengal test (RBT) and complement fixation test (CFT) results on the serums from 2,307 cattle (from herds where infection was still present after a minimum of 3 complete herd tests) showed that 327 were positive in the CFT and 246 were positive in the RBT (p less than 0.001). Subsequent testing by the RBT, CFT and the indirect haemolysis test (IHLT) of 722 serums from cattle slaughtered as part of infected herds showed that of 177 cattle positive on culture, 138 were positive in all 3 tests, 9 were negative in all 3 tests and no animal positive on culture had a reaction only in the RBT. In the 177 cattle from which B. abortus was isolated, positive reactions in the CFT occurred in the serums of 159 of them. Application of the RBT as a screening test followed by a confirmatory CFT would have resulted in 149 of the 177 cattle being positive and application of the CFT/IHLT (double test) on the serums of all cattle in the herds would have resulted in 168 or the 177 being regarded as positive.     

湖南高致病性猪蓝耳病隐性感染情况调查

用ELISA和RT-PCR的方法对采集自湖南省内20个规模场1007份血清和3个市级定点屠宰场50份猪肺门淋巴结进行蓝耳病血清抗体检测和高致病性猪蓝耳病病毒的检测。结果1007份血清中,蓝耳病抗体阳性率为72.9%(734/1007),高致病性猪蓝耳病病毒携毒率为3.2%(32/1007),50份肺门淋巴结病毒阳性率为16%(8/50),其中,蓝耳病免疫与非免疫猪血清其抗体阳性率相差不显著,种猪的抗体阳性率明显高于商品猪,而其病毒携毒率为0%。部分规模猪场和眼观健康的育肥猪存在高致病性猪蓝耳病病毒的隐性感染。     

携带传染性鼻气管炎病毒牦牛肝功能指标检测

采用生物化学方法对34份携带传染性鼻气管炎病毒（IBRV）牦牛（阳性组）血清和18份健康牦牛（阴性组）血清的肝功能指标进行了测定。 结果表明: 阳性组血清的总蛋白、球蛋白、β 球蛋白显著低于阴性组（P<0.05）;而AST、ALT及AST/ALT 均高于阴性组血清,差异极显著（P<0.01）,且部分阳性牛的肝脏损害严重（11.77%）。     

A microplate enzyme immunoassay for detecting and measuring antibodies to Babesia bovis in cattle serum

A microplate enzyme immunoassay (EIA) is described for measuring IgG antibody to Babesia bovis in cattle serum. B. Bovis antibody status (whether positive or negative) and the amount of B. Bovis antibody (EIA score), were measured by comparison with reference serums. The EIA was shown to be specific for B. Bovis, and EIA score correlated well with EIA titre. Comparison of EIA with the Indirect Fluorescent Antibody Test (IFAT) showed more than 95% agreement between the methods and disagreement in only 1.6% of serum samples tested. The remaining 3.2% were positive by EIA and suspected positive by IFAT. The EIA was shown, by titrating positive serums, to be more sensitive than IFAT, which explained its tendency to detect more positive serums than IFAT. EIA detected B. bovis antibody in experimentally infected cattle by day 14 post infection (pi) and for at least 268 days pi. EIA score for B. bovis antibody in immune cattle increased significantly (p less than 0.05) following heterologous strain challenge.     

A COMPLEMENT-FIXATION TEST FOR ENZOOTIC PNEUMONIA OF PIGS USING A COMPLEMENT DILUTION METHOD

Complement-fixing antibody to Mycoplasma hyopneumoniae in the serums of pigs experimentally infected with enzootic pneumonia was demonstrated by comparing the haemolytic titre of guinea-pig complement titrated in the presence of heated test serum, M. hyopneumoniae antigen and unheated normal pig serum with the titre obtained when the antigen was omitted. The haemolytic titres against sensitised sheep erythrocytes were determined after a fixation period of 16 to 18 hours at 5°C. When serums, collected at intervals of 3 to 7 days, from 43 pigs exposed to pigs experimentally infected with enzootic pneumonia were tested, 4.6 or more complement units were first fixed 14 to 44 (mean 23.4) days after contact began. Serums collected subsequently fixed from 4.6 to more than 31 complement units. This positive reaction usually persisted until the pigs were killed 4 to 35 weeks after contact began. Thirty-three had gross enzootic pneumonia lesions and 9 had lung lesions detected microscopically. Serum antibody was not detected in 73 weaned pigs aged 7 weeks in a pneumonia-free herd but serums from 9 of 15 unweaned piglets aged 9 to 14 days in the same herd, fixed between 3 and 7 complement units.     

The prevalence and distribution of leptospiral titres in cattle and pigs in Queensland

Serological test results for leptospiral species on serums from cattle and pigs performed by the diagnostic laboratories of the Queensland Department of Primary Industries from July 1973 to June 1976 were used to determine the prevalence and geographical distribution of 3 leptospiral serotypes in Queensland. There was a higher prevalence of antibodies to L. hardjo than to L. pomona in cattle, whereas in pigs the prevalence of antibodies to L. pomona was much higher than that for L. tarassovi or L. hardjo. Feral pigs had a particularly high prevalence of L. pomona antibodies. There is a different geographical distribution of antibodies to L. pomona and L. hardjo. L. hardjo antibodies appear to be fairly uniformly distributed but there is a higher prevalence of L. pomona antibodies in low rainfall areas. This relationship was shown to be significantly correlated.