斑点金标免疫渗滤法快速检测牛血吸虫病抗体的效果观察

目的观察斑点金标免疫渗滤法检测牛血吸虫病抗体的效果。方法使用斑点金标免疫渗滤诊断试剂盒检测粪检血吸虫病牛血清(血纸)146头份,疫区耕牛血清(血纸)562头份,并用间接血凝法(indirect haem aggluti-nation assay,IHA)法作对比。结果146头份粪检阳性血清(血纸)检测为阳性者140头份,阳性检出率95.9%,IHA法检测为阳性者141头份,阳性检出率96.6%。结论斑点金标免疫渗滤法检测牛血吸虫病具有快速、简便、灵敏度高、特异性强等优点,但仍有一定漏检,有待于进一步研究提高。     

斑点金标免疫渗滤新技术检测家畜血吸虫病的效果试验

采用血吸虫病斑点金标免疫渗滤诊断技术在永胜县做了动物血吸虫病感染符合率、特异性试验,共检测2 800份牛血纸,结果检出阳性78头;检测755份牛血清,检出阳性72头;再对被检查的2 800头牛采用粪便毛蚴孵化法检出阳性74头。免疫学诊断结果与病原检查结果阳性符合率血纸达94.8%,血清达97.3%。同时在县内非疫区乡(镇)做了阴性符合率试验,符合率达100%。血吸虫病斑点金标免疫渗滤诊断技术在永胜县6个血防区试验运用,取得了成功。     

检测家畜血吸虫抗体的二步金标免疫渗滤法研究

将待检家畜血纸浸出液点在硝酸纤维素膜上，以金标记的血吸虫虫卵可溶性抗原为探针（以下简称血吸虫抗原胶体金），建立了检测家畜血吸虫抗体的二步金标免疫渗滤法（Two-step Dot Immunogold Filtration Assay，T—DIGFA）。家畜血吸虫病阳性血样在50-90S形成肉眼可观察的红色斑点，可进行快速诊断。该法可以检测出人工感染血吸虫尾蚴7d和7d以上的阳性牛和兔血纸抗体，与肝片吸虫、锥虫、蛔虫病之间无交叉反应。T—DIGFA检测粪孵血吸虫毛蚴阳性牛血纸139份、阴性牛血纸130份，与粪孵法阳性符合率为100％，阴性符合率为99．2％；T—DIGFA与斑点酶联SPA法阳性符合率和阴性符合率完全相同。血吸虫抗原胶体金在4～8℃保存期可达6个月、室温保存期为3个月。试验证实，T-DIGFA有很高的敏感性、特异性、重复性和稳定性，适合于基层单位和现场进行家畜血吸虫病抗体的快速诊断、普查和检疫。     

粪孵法与斑点酶标法检测牛血吸虫病浅析

通过粪孵法与斑点酶标法对疫区牛进行血吸虫师检测对比试验表明，用斑点酶标法检测，其操作简便易行快速，不受环境影响，工效是粪孵法的１０倍，并且每头被测牛节省诊断费２．２元。     

血纸间接血凝与斑点金标诊断血吸虫病效果对比试验

为探讨快速、准确、简便、经济的血吸虫病血清学诊断方法,减轻劳动强度,作者于2007年6月在常德市贺家山农场对290份血样(其中牛190份,羊100份)进行了血纸间接血凝诊断与斑点金标诊断效果对比试验。结果表明:斑点金标诊断法操作简单,所需检测器械较少,检测时间短,但成本相对较高;间接血凝法操作较为复杂,检测时间较长,但成本相对较低。     

应用斑点酶标法诊断耕牛血吸虫病

应用斑点酶标法诊断耕牛血吸虫病四川省兽医总站（６１００４１）毛光琼谢智明阳爱国德阳市兽医站朱庭荣什邡市畜牧局吴兴国目前，全省各地均采用粪孵法诊断耕牛血吸虫病。但由于该方法劳动强度大、时间长、检出率低，一般情况下，每天每人只能检测３０头份粪便，检出率...     

斑点金免疫渗滤测定法检测耕牛血吸虫病的效果观察

免疫胶体金技术是以胶体作为示踪标志物应用于抗原抗体的一种新型免疫标记技术，没有潜在致癌物质的酶显色底物。斑点金免疫渗滤测定法是将斑点酶标与免疫胶体金结合起来的一种新型免疫标记技术。该测定方法是将被检抗体（血清或血纸浸出液）直接点样在硝酸纤维素膜上，滴加胶体金标记的血吸虫抗原，特异性抗原与抗体很快结合，在反应处发生金颗粒聚集，1min左右形成肉眼可见的红色斑点，判定检测效果。     

斑点金标免疫渗滤新技术检测家畜血吸虫病的效果试验

斑点金标免疫渗滤新技术(简称金标法)检测家畜血吸虫病抗体,在本省进行了特异性、敏感性、符合率等试验.金标法能检出人工感染犬血吸虫14 d后的血吸虫病抗体,检出率为100%,可用于血吸虫病早期诊断.粪孵100头份血吸虫病阳性的牛血纸,用金标法检测全部为阳性,阳性符合率为100%;在非疫区的2个县分别采牛血纸100份,用金标法进行检测,未查出血吸虫病阳性牛,阴性符合率为100%.在金标法检测4 000份牛血纸,检出阳性牛为129头,再对这129头牛采粪孵化,有血吸虫毛蚴的牛128头,金标法与病原学检查法阳性符合率为99.2%(99%～100%).在本省54个血防疫区县推广应用,取得了显著的社会经济效益.     

斑点金免疫渗滤测定法检测耕牛血吸虫病

免疫胶体金技术是以胶体金作为示踪标志物应用于抗原抗体的一种新型的免疫标记技术，没有潜在致癌物质的酶显色底物。斑点金免疫渗滤测定法是将斑点酶标与免疫胶体金结合起来的一种新型的免疫标记技术。将被检抗体（血清或血纸浸出液）直接点样在硝酸纤维素膜上，滴加胶体金标记的血吸虫抗原，特异性抗原与抗体很快结合，在反应处发生金颗粒聚集，1min左右形成肉眼可见的红色斑点。     

斑点金标免疫渗滤法诊断家畜血吸虫病的效果

我县从2005-2006年推广应用斑点金标免疫渗滤法(DIGFA)诊断家畜血吸虫病新技术,该项新技术具有快速、简便、准确、省时、省力等优点,使阳性病畜得到及时治疗,取得了满意的效果。1材料与方法1.1诊断试剂DIGFA诊断试剂及Dot-ELISA诊断试剂均由浙江省农业科学院畜牧兽医研究所提供     

均相光激化学发光免疫分析技术的研究进展

均相光激化学发光免疫分析技术是一种基于纳米微珠的化学发光的新型技术,其具有更高的敏感度、均一性、背景低,且不用洗涤及样本需求量少等特点。该技术可用于生物标志物、激酶及抗原抗体的检测,蛋白：蛋白相互作用的检测,高通量分析的研究及疾病诊断等方面。优化和发展均相光激化学发光免疫分析技术将会在更多研究领域中得到应用。     

Comparative titers of egg assay against immunofluorescent assay of Chlamydia psittaci.

A comparison of titers was made between an egg assay and a direct fluorescent antibody assay of three chlamydial strains propagated in Vero cells with and without cortisone plus cytochalasin B. The titer of NJ-1 strain was similar in the egg titration and the fluorescent antibody assay in the untreated sample and a little lower for the sample treated with cytochalasin B and cortisone. The SCT and CDC strains had approximately the same titers in the egg titration and the fluorescent antibody assay for samples with and without the antimetabolites.     

饲用纤维素酶测定方法的探讨

本文在总结纤维素酶作用机理的基础上，根据饲料中所含不溶性纤维素的特性及其降解时对酶系要求，推荐以滤纸为底物测定饲用纤维素酶活力。     

家禽饲料代谢能的评定技术

家禽饲料代谢能的评定是家禽饲料营养价值评定中一个极其重要的内容，是研究家禽营养需要量的基础，也是配制最佳饲料配方的关键，对提高家禽的生产效率、节约饲料资源具有重要意义。因此，探索家禽饲料代谢能的快速、简便、准确的评定方法一直是动物营养研究中的热点。世界各国的营养学家们经过几十年的探索和努力，摸索出了许多科学、实用的代谢能测定方法，将这些方法归类主要有常规法、快速测定法和体外法等。     

Plaque assay of bovine rotavirus

Incorporation of trypsin and diethylaminoethyl-dextran in the overlay was found to be necessary for infectivity assay of the UK strain of bovine rotavirus by plaque assays. Small plaques of about 1 mm in radius were formed in BGM cells. Large plaques of about 3–4 mm in radius were consistently produced in monolayers of secondary calf kidney cultures.Key words: bovine rotavirus, plaque assay, trypsin, diethyl-aminoethy1-dextran     

Validation of a von Willebrand factor antigen enzyme-linked immunosorbent assay and newly developed collagen-binding assay

No single test is comprehensive enough to detect all of the variants of von Willebrand Disease (VWD), making determination of both concentration and function of von Willebrand Factor (VWF) important for an accurate diagnosis. The objective of the study was to validate a newly developed VWF collagen binding assay (VWF:CB) and VWF antigen enzyme-linked immunosorbent assay (ELISA) developed at the Ontario Veterinary College (OVC VWF:Ag). Linearity, sensitivity, and coefficients of variation were determined. The Asserachrom VWF:Ag ELISA was used as the reference assay for this study. Concordance correlation and Bland-Altman plots were used to evaluate agreement between both VWF:Ag assays. The VWF:CB accuracy was assessed by degree of association with the VWF:Ag assays, and the VWF:Ag to VWF:CB ratio. All assays were assessed for their ability to distinguish between VWD negative and VWD positive patients. Linearity, intra-assay coefficients of variation, and inter-assay coefficients of variation were acceptable for both the newly developed VWF:CB (R² = 0.97, average CV = 4.4, and 15, respectively) and OVC VWF:Ag assays (R² = 0.96, average CV = 7.9, and 5.9, respectively). Agreement between the OVC VWF:Ag assay and reference assay was excellent (ρ_c = 0.89), and although differences between assay results precluded interchangeable use of the assays, both successfully distinguished VWD positive and VWD negative dogs (P < 0.0001). The VWF:CB showed a strong association with both VWF:Ag assays (R² = 0.86, 0.82) and VWF:Ag to VWF:CB ratios (≤ 1) were as expected. The excellent performance of both assays in this validation study confirm their reliability and potential for clinical application.     

H3N2亚型犬流感病毒血凝及血凝抑制试验的研究

为优化H3N2亚型犬流感病毒(CIV)的血凝抑制(HI)试验方法,本研究应用不同种类红细胞进行CIV的血凝(HA)试验,应用不同种类红细胞和不同血清处理方法进行CIV的HI试验,评价其对HA和HI试验的影响。结果表明,H3N2亚型CIV对鸡和犬红细胞的凝集性最好,对小鼠、猪和牛红细胞的凝集性较差。HI试验应用鸡红细胞悬液效果最好,受体破坏酶(RDE)和高碘酸钾处理可以有效去除犬血清中非特异血凝抑制素,但高碘酸钾对血清特异性抗体有损耗。本研究筛选出H3N2亚型CIV HI试验的最佳方法,为犬流感的血清学诊断提供技术支持。     

Comparison of dot immunobinding assay, enzyme-linked immunosorbent assay and immunodiffusion for serodiagnosis of paratuberculosis.

A rapid, simple and inexpensive dot immunobinding assay (DIA) was evaluated for the serodiagnosis of paratuberculosis in cattle. The assay was performed on nitrocellulose strips which were dotted with purified protoplasmic antigen of Mycobacterium paratuberculosis. After incubation with test serum samples, the bound antibodies were detected using an enzyme-amplified immunostaining procedure. The efficacy of DIA as a screening test for paratuberculosis was compared to that of an enzyme-linked immunosorbent assay (ELISA), a modified agar gel immunodiffusion (mAGID) test, and an AGID test using 329 serum samples from cattle which were examined for M. paratuberculosis infection by a sensitive fecal culture technique. The DIA and ELISA had comparable results and both of the enzyme immunoassays had higher sensitivity than tests based on AGID. The sensitivity of all four tests was influenced by the intensity of fecal bacterial shedding. Preabsorption of sera with Mycobacterium phlei increased the sensitivity of both enzyme immunoassays. the specificity but reduced the sensitivity of both enzyme immunoassays.