蓝舌病灭活疫苗免疫羊ELISA和琼扩监测比较试验

用19只绵羊（注苗前BTV琼扩和ELISA均为阴性）注射BTV灭活苗后7～139天的血清进行BTVELISA和琼扩平行试验，结果表明，注苗后7～30天11只试验丰ELISA阳性，注苗40天13只羊ELISA阳性，注苗90天以后全部试验羊只均为ELISA阳性，而琼扩试验则始终为阴性结果。     

Sensitivity and specificity of two serological tests for the detection of ovine paratuberculosis

OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA.     

小鼠抗重组山羊胎盘催乳素多克隆抗体的制备及其在山羊胎盘组织中定位的研究

用rcPL对6-8周龄的昆明小鼠进行免疫注射制备多抗,第3次加强免疫后第6天采集少许血清,用琼脂凝胶免疫扩散（AGID）和间接酶联免疫吸附试验（ELISA）检测抗体的产生情况;分娩后立即通过剖腹产的方法获得山羊胎盘组织,用获得的抗血清通过免疫组织化学（IHC）和间接荧光抗体试验（IFA）的方法定位胎盘催乳素。结果表明,琼脂扩散检测抗原浓度为100μg／mL,用ELISA中酶标仪检测450nm处OD值,免疫血清与阴性对照的比值P／N为7．77;IHC和IFA显示胎盘催乳素在肉阜和子叶的双核细胞及单核细胞中表达。使用背部皮下多点注射的方法对昆明小白鼠进行免疫来制备重组山羊胎盘催乳素多克隆抗体是可行的,制备的抗体能满足免疫组织化学和荧光免疫组织化学检测胎盘催乳素的要求。     

禽流感病毒重组核蛋白在琼扩诊断中的应用

利用杆状病毒表达禽流感病毒型特异性抗原－核蛋白,并就重组核蛋白作为禽流感琼扩诊断抗原地特异性和敏感性及作为抗原的其它特性进行了研究,并与普通全病毒琼扩抗原进行了应用比较试验,对３５４份现地可疑血清样品进行了检测,结果表明：该抗原可以特异性地定性检测出所有１５个禽流感病毒亚型的抗血清,与普通全病毒琼扩抗原的检测结果一致,但沉淀线更细密、清晰,保存方便;与我国广泛流行的１５种禽类其它病原体的抗血清没有任何交叉反应,表明该重组核蛋白作为禽流感琼扩抗原特异、敏感、生物安全性更可靠和生产成本更低廉,有利于进一步扩大禽流感疫情调查范围和加大监测力度。     

接种传染性法氏囊病基因工程重组亚单位油乳剂疫苗的产蛋鸡血清抗体与蛋黄抗体的相关性分析

用笔者研制的传染性法氏囊病(IBD)基因工程重组亚单位油乳剂疫苗免疫产蛋鸡,接种后第22d及以后每周进行一次无菌采血,分离血清,用琼脂免疫扩散试验(AGID)检测IBD血清抗体水平;每周用同样的方法检测蛋黄中IBD抗体水平。对血清抗体水平和蛋黄中抗体水平进行相关性统计分析,得出两者之间的相关系数,并进行显著性检验。统计分析结果表明,相关系数r=0.7782,表明血清抗体水平和蛋黄中抗体水平相关关系属于强正直线相关;t值为14.02,t>t(∞)=2.576,差异极显著(P<0.01),即血清抗体水平和蛋黄中抗体水平之间的相关系数极显著;F值为190.55,F>F0.01(1,128)=6.84,差异极显著(P<0.01),即血清抗体水平和蛋黄中抗体水平之间的相关关系极显著。     

实验感染OPPV绵羊血清中抗体水平动态检测

应用建立的ＥＬＩＳＡ方法和常规的ＡＧＩＤ方法对人工感染ＯＰＰＶ的试验羊进行抗体水平动态检测，结果表明，ＥＬＩＳＡ敏感性高，特异性好。通过颈静脉途径接种２个月后，可出现ＥＬＩＳＡ阳性反应（３／５），而通过气管接种为１个月（２／２）；相对来讲ＡＧＩＤ阳性出现的时间要迟２￣５个月，很难检出低水平的抗体。     

传染性法氏囊病免疫保护试验中3种评定指标的比较

100只SPF鸡均分为5组，A、B、c3组免疫后攻毒，接种3批次自制的IBD基因工程重组亚单位油乳剂疫苗；D组不免疫不攻毒，E组不免疫而攻毒。免疫后第22天，感染IBDV强毒株BC-6／85。攻毒后第4天，将所有存活的鸡只以颈脱臼致死，收集法氏囊，以3种方法和指标（法氏囊眼观病变；法氏囊显微病变；法氏囊中IBDV抗原检测）进行分析，以评定免疫保护率。结果显示，以这3种方法和指标评定，A组免疫保护率为90％，B、C组免疫保护率均为95％；试验鸡A3、A14、B7、C16、E1～E20，法氏囊眼观病变明显，法氏囊显微病理损伤评分为3～5分，琼脂免疫扩散试验检测法氏囊中IBDV抗原均为阳性；其他试验鸡，法氏囊眼观无明显异常，法氏囊显微病理损伤评分在3分以下，琼脂免疫扩散试验检测法氏囊中IBDV抗原均为阴性。结果表明，这3种方法和指标在IBD疫苗免疫保护试验评定中具有很好的一致性。     

Serodiagnosis of equine infectious anemia by agar gel immunodiffusion and ELISA using a recombinant p26 viral protein expressed in Escherichia coli as antigen

We used a p26 recombinant protein (p26r) from equine infectious-anemia virus (EIAV) expressed in Escherichia coli as antigen to standardize an agar-gel immunodiffusion (AGIDp26r) test and an indirect ELISA (ELISAp26r) for the detection of antibodies against EIAV in 720 equine sera from Brazil. We evaluated the tests's relative diagnostic sensitivities (relSe) and relative diagnostic specificities (relSp) against a commercial AGID kit (Idexx™, USA). We used three sera panels: panel A—196 AGID-negative sera from an AIE non-endemic controlled area; panel B—194 AGID-negative sera from an AIE endemic area and panel C—330 AGID-positive sera from an AIE endemic area. ELISAp26r cut-off value was defined with TG-ROC using sera from panels A and C. AGIDp26r showed an agreement of 100% with the commercial kit. When applied to sera from panels A and C, ELISAp26r showed an agreement of 100% with the kit, but, although relSe was 100% for panel C, the ELISAp26r had relSp of 93.3%.