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1.
本文建立的Ｄｏｔ－ＥＬＩＳＡ对猪衣原体ＩｇＧ的最小检测量为１．３３×１０－９ｇ．与间接补体结合试验（ＩＣＦ）比较；检测１１０头份猪血清样品，Ｄｏｔ－ＥＬＩＳＡ检出５２份（４７．２７２％，效价≥６４），ＩＣＦ法检出３２份（２９．０９％，效价≥８）．用异种动物衣原体标准阳性血清作阻断试验，证明Ｄｏｔ－ＥＬＩＳＡ特异性好（阻断可使原效价下降至少８个单位以上），诊断膜与猪瘟阳性血清、猪水疱病阳性血清、猪口蹄疫阳性血清、猪弓形体阳性血清不出现有意义的交叉反应（效价≤１６）．诊断膜片置室温（２５℃左右）、４℃和－２０℃保存１个月后反应效果不变，对照反应均成立，重复性好（重复符合率为９０．９％），反应效果不受不同批次微量反应板的影响，反应板可重复使用多次，试剂用量较常规ＥＬＩＳＡ大大节省，各步骤均可做到严格定量，操作简便（３小时内可完成），不需要特殊检测仪器，结果客观，肉眼易于判断，是微生物和传染病及寄生虫病诊断技术标准化很有前途的新技术之一． 相似文献
2.
银加强胶体金技术检测猪细小病毒的研究 总被引：18，自引：1，他引：17
本研究首次成功地建立了检测猪细小病毒（ＰＰＶ）抗原的银加强胶体金检测法（ＳＥＣＧＡ），并确定了操作流程中的最佳试验条件。应用该法对纯化ＰＰＶ抗原的最低检出量为０．３１２５ｎｇ／点，其敏感性分别为间接Ｄｏｔ－ＥＬＩＳＡ和ＨＡ的８倍和１０００倍。特异性阻断试验和交叉反应试验证明ＳＥＣＧＡ具有较高的特异性。２０份样本ＳＥＣＧＡ的检测结果与病毒分离和鉴定结果完全相符。ＳＥＣＧＡ和间接Ｄｏｔ－ＥＬＩＳＡ对７７份样本检测的阳性率分别为８３．１％和８０．５％，其阳性符合率为９６．９％。研究结果表明本法具有经济、敏感性、特异性强等优点，可用于ＰＰＶ感染的特异诊断。为胶体金技术应用于畜禽传染病的诊断和研究奠定基础。 相似文献
3.
早期诊断与治疗猪囊虫病试验 总被引：1，自引：0，他引：1
为探索早期诊断与治疗猪囊虫病的有效方法，采用ＥＬＩＳＡ诊断大通地区的猪血样１３８２头份，并对其中的７７头进行了剖检验证，还用丙硫咪唑治疗已确诊患囊虫病的猪５８头。试验结果：用ＥＬＩＳＡ检出猪囊虫阳性率为８．０３％；ＥＬＩＳＡ血检与剖检结果的阳性符合率为９２．３％，阴性符合率为９５．３％，总体符合率为９４．８％；丙硫咪唑对猪囊虫的杀灭率为９８．７％，治愈率为９３．７％，经治疗后让机体自身修复７—９个月时，达到鲜销标准的猪为７２．８％—９０．９％。 相似文献
4.
斑点酶联免疫吸附试验检测猪繁殖与呼吸综合征病毒抗体方法的建立 总被引：6，自引：0，他引：6
首次建立了斑点－酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）检测猪繁殖与呼吸综合征病毒（ＰＲＲＳＶ，美洲株）抗体的方法。特异性鉴定表明，ＰＲＲＳＶ不与猪瘟病毒、猪伪狂犬病病毒、猪细小病毒阳性血清反应；与美国进口的ＰＲＲＳＶ抗体ＥＬＩＳＡ诊断试验盒检测结果比较，３５份猪血清的阴、阳性检出总符合率为８２．９％（２９／３５），其中Ｄｏｔ－ＥＬＩＳＡ的阳性检出率略高于进口试剂盒的检出率。 相似文献
5.
采用四种方法：即①单克隆抗体检测抗原的酶联免疫吸附试验（Ａｇ－ＥＬＩＳＡ）②检测抗体的酶联免疫吸附试验（Ａｂ－ＥＵＳＡ）③表膜检查（ＢＣＥ）④小白鼠接种（ＭＩ），检查了肯尼亚１８３头骆驼锥虫的循环抗原，ＢＣＥ查出３７头阳性２０％），ＭＩ６０头阳性（３３％），Ａｇ－ＥＬＩＳＡ６３头阳性（３４％），Ａｂ－ＥＬＩＳＡ检出９０头阳性（４９％）。其中ＢＣＥ未能检出的２４头中，Ａｇ－ＥＬＩＳＡ检出１８头（７５％）。所有阳性头数中，Ａｇ－ＥＬＩＳＡ检出９３％，Ａｂ－ＥＬＩＳＡ９５％。根据５５头的结果，阳性与阴性之间Ａｇ－ＥＬＩＳＡＯＤ值差异极显著（Ｐ＜０．０００１），Ａｂ－ＥＬＩＳＡＯＤ值差异不显著。因此，用Ａｇ－ＥＬＩＳＡ或结合ＢＣＥ诊断伊氏锥虫感染比用ＡＢ－ＥＬＩＳＡ更理想。 相似文献
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应用ＳｅｐｈａｄｃｘＧ－２００层析法纯化鸡减蛋综合症病毒，利用ＮＣ膜作为载体，成功建立了检测ＥＤＳ－７６血请抗体的斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）。抗原包被浓度为２μｇ／ｍｌ，被检血清浓度为１：２０，酶标兔抗鸡１ｇＧ浓度为１：２００．底物溶液最适ｐＨ值为７．２。对Ａ－Ｆ６个养禽场随机抽检血清１６０份，分别用Ｄｏｔ－ＥＬＩＳＡ、ＨＩ和ＡＧＰ检测，Ｄｏｔ－ＥＬＩＳＡ检出阳性率为５１．９％，ＨＩ检出阳性率为４６．９％，ＡＧＰ检出阳性率为３１．９％。对１４０日龄鸡人工感染试验，测定抗体消长规律。本方法不需特殊仪器。适用于基层兽医部门和养鸡场对ＥＤＳ－７６的血清学诊断和流行病学调查。 相似文献
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李国清 《国外兽医学(畜禽疾病)》1994,15(1):40-43
采用四种方法：即①单克隆抗体检抗原的酶联免疫吸附试验（Ａｇ－ＥＬＩＳＡ）②检测抗体的在酶联免疫吸附试验（Ａｂ－ＥＬＩＳＡ）③表膜检查（ＢＣＥ）④小白鼠接种（ＭＩ），检查了肯尼亚１８３头骆驼锥虫的循环抗原，ＢＣＥ查出尼亚１８３头骆驼锥早的循环抗原，ＢＣＥ查出３７头阳性２０％），ＭＩ６０头阳性（３３％），Ａｇ－ＥＬＩＳＡ６３头阳性（４９％），Ａｂ－ＥＬＩＳＡ检出出的２４头中，Ａｇ－ＥＬＩＳＡ检出１８头 相似文献
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ELISA法检测犬腹泻粪样中的犬冠状病毒 总被引：2，自引：0，他引：2
用ＦＥ细胞增殖犬冠状病毒（ＣＣＶ）参考株，分别免疫家兔和ＢＡＬＢ／ｃ小鼠制备ＣＣＶ多抗和单抗，建立了夹心ＥＬＩＳＡ及Ｄｏｔ－ＥＬＩＳＡ诊断方法。在检测的８４例犬腹泻粪样中，多抗、单抗夹心法显示ＣＣＶ阳性１６例，Ｄｏｔ－ＥＬＩＳＡ阳性１３例，后１３例包括在前１６例中。从８４例腹泻犬粪样中随机取３８例作ＣＣＶ、犬细小病毒（ＣＰＶ）双项检测，ＣＣＶ阳性１６例，ＣＰＶ阳性６例，ＣＣＶ、ＣＰＶ混合感染４例。结果显示，在南京地区流行的犬腹泻中，ＣＣＶ感染比例有超过ＣＰＶ的趋势。 相似文献
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ELISAA法检测犬腹泻粪样中的犬冠状病毒 总被引：6，自引：0，他引：6
用ＦＥ细胞增殖犬冠状病毒（ＣＣＶ）参考株，分别免疫家兔和ＢＡＬＢ／ｃ小鼠制备ＣＣＶ多抗和单抗，建立了夹心ＥＬＩＳＡ及Ｄｏｔ－ＥＬＩＳＡ诊断方法。在检测的８４例犬腹泻粪样中，多抗、单抗夹心法显示ＣＣＶ阳性１６例，Ｄｏｔ－ＥＬＩＳＡ阳性１３便，后１３例包括在前１６例中，从８４例腹泻犬粪样中随机取３例作ＣＣＶ、犬细小病毒（ＣＰＶ）双项检测，ＣＣＶ阳性１６例，ＣＰＶ阳性６例，ＣＣＶ、ＣＰＶ混合感染４例。结 相似文献
10.
非洲猪瘟间接ELISA诊断试剂盒的研究 总被引：12，自引：6，他引：6
用带有编码非洲猪瘟病毒衣壳蛋白Ｐ７２ 基因的重组杆状病毒（Ｂａｃｐ７２）作载体,在ｓｆ９细胞中表达并得到重组Ｐ７２蛋白,ＳＤＳ－ＰＡＧＥ可得到分子量在 ７２ｋＤａ左右的电泳带。用标准阳性非洲猪瘟血清对 Ｐ７２蛋白进行 ＥＬＩＳＡ检测,证明该蛋白具有生物学活性。用Ｐ７２作为间接法的包被抗原,对ＥＬＩＳＡ反应条件进行了优化。确定最佳包被液为ＰＢＳ（ｐＨ７．２）、最佳封闭液为１％ＰＣＴ、最佳血清稀释液为４％ＰＥＧ６０００／ＰＢＳ、最佳冲洗液为０．５Ｍ ＮａＣｌ／０．５％Ｔｗｅｅｎ－２０／ＰＢＳ（ｐＨ７．２）。本实验反应体系采用５０μｌ的微量法,可节约试剂及抗原。反应在２小时内即可完成,达到了快速诊断的目的。包被了抗原并用封闭液封闭后的酶标板密封后保存于－２０℃的冰箱中,至少可以保存５个月。阻断试验和交叉试验表明ＥＬＩＳＡ法有良好的特异性。间接ＥＬＩＳＡ比Ｄｏｔ－ＥＬＩＳＡ法具有更高的灵敏性。血清学调查没有得到阳性结果,与我国实际情况相符。用Ｂａｃｐ７２表达的非洲猪瘟病毒Ｐ７２蛋白抗原作为间接ＥＬＩＳＡ的检测抗原来检测非洲猪瘟血清具有快速、简单、无感染的特点。本实验为非洲猪瘟ＥＬＩＳＡ检测试剂盒的最终组装提供了实验依据。 相似文献
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T Miyamae 《American journal of veterinary research》1980,41(4):584-585
Pancreas-passaged avian encephalomyelitis (AE) virus was transmitted horizontally in a group of 40 (1-day-old) chicks within 3 weeks after they were intermingled with two orally infected 1-day-old chicks. Viral antigen was detected in the pancreas of these contact-exposed chicks. After 5 weeks, contact-exposed chicks developed high titers against AE virus, but the chicks did not develop clinical signs of AE. The passaged virus could not be recovered from feces of six immunized chicks. 相似文献
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三批禽脑脊髓炎(AE)油佐剂灭活疫苗免疫开产前种鸡,种鸡免疫2~3个月后所产种蛋对禽脑脊髓炎病毒(AEV)鸡胚易感性试验的受保护率达100%,而同期所产种蛋孵出的仔代雏鸡,在出雏后3周内对AEV VR株攻毒的受保护率亦达100%; 种鸡免疫6~7个月后所产种蛋对AEV VR鸡胚易感性试验的受保护率达75%~80%,而同期所产种蛋孵出的仔代雏鸡,在出雏后3周内对AEVVR株攻毒的受保护率达70%~90%。试验结果表明,AE灭活疫苗AEV鸡胚易感性试验的鸡胚保护率与同期雏鸡对AEV VR肌肉途径攻毒的保护率基本相同。 相似文献
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Several subgroup J-like avian leukosis viruses (ALV-Js) were isolated from broiler breeder (BB) and commercial broiler flocks experiencing myeloid leukosis (ML) at 4 wk of age or older. In all cases, diagnosis of ML was based on the presence of typical gross and microscopic lesions in affected tissues. The isolates were classified as ALV-J by 1) their ability to propagate in chicken embryo fibroblasts (CEF) that are resistant to avian leukosis virus (ALV) subgroups A and E (C/AE) and 2) positive reaction in a polymerase chain reaction with primers specific for ALV-J. The prototype strain of these isolates, an isolate termed ADOL-Hc1, was obtained from an adult BB flock that had a history of ML. The ADOL-Hc1 was isolated and propagated on C/AE CEF and was distinct antigenically from ALV of subgroups A, B, C, D, and E, as determined by virus neutralization tests. Antibody to ADOL-Hc1 neutralized strain HPRS-103, the prototype of ALV-J isolated from meat-type chickens in the United Kingdom, but antibody to HPRS-103 did not neutralize strain ADOL-Hc1. On the basis of both viremia and antibody, prevalence of ALV-J infection in affected flocks was as high as 87%. Viremia in day-old chicks of three different hatches from a BB flock naturally infected with ALV-J varied from 4% to 25%; in two of the three hatches, 100% of chicks that tested negative for virus at hatch had evidence of viremia by 8 wk of age. The data document the isolation of ALV-J from meat-type chickens experiencing ML as young as 4 wk of age. The data also suggest that strain ADOL-Hc1 is antigenically related, but not identical, to strain HPRS-103 and that contact transmission of ALV-J is efficient and can lead to tolerant infection. 相似文献
14.
Application of the Fluorescent Antibody Technique to the Diagnosis of Avian Encephalomyelitis 总被引：3，自引：3，他引：0 
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下载免费PDF全文 P. R. Ide 《Canadian journal of veterinary research》1974,38(1):49-55
A direct fluorescent antibody test was applied to the diagnosis of avian encephalomyeltis in experimental chicks and in chicks from three natural outbreaks of the disease. Intracerebrally inoculated chicks exhibited maximum fluorescence in brain tissue during the early clinical stages of the disease and similar observations were made on natural cases. Best fluorescence was obtained with frozen sections of cerebellum and pancreas from chicks approximately three weeks of age, but after four weeks of age it was not possible to diagnose the naturally-occurring (field) disease by this method. In these cases a diagnosis of avian encephalomyelitis was confirmed by the application of the fluorescent technique to the brains of either (a) chicks which had hatched from eggs inoculated with brain homogenates from the field cases or (b) hatched chicks which had been inoculated with the homogenates by the intracerebral route. 相似文献
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Molly E. Church Sridhar M. Veluvolu Amy C. Durham Kevin D. Woolard 《Veterinary and comparative oncology》2019,17(4):578-584
Olfactory neuroblastoma (ONB) is a rare intranasal neoplasm in both dogs and humans. Similar clinical presentation and overlapping histologic and immunohistochemical features of ONB with other intranasal neoplasms can make diagnosis and treatment of intranasal neoplasia challenging. Furthermore, in part because of their rarity, there is a lack of reporting on therapeutic regimen for these neoplasms. In humans, initial debulking surgery is usually followed by radiation therapy. Here we report on the histologic, immunohistochemical, and ultrastructural characteristics of canine ONB and report on the clinical progression of cases treated with radiation therapy. In all nine canine ONB examined here, neoplastic cells were arranged in a lobular manner amidst a prominent neurofibrillary matrix and had features consistent with Grade III (high grade) ONB. The neoplastic cells demonstrated positive immunohistochemical staining for TuJ‐1, a Class III beta‐tubulin neuronal cytoskeletal protein, and variable staining for other markers, including chromogranin, synaptophysin, AE1/AE3 and MAP2. The longest surviving case was treated with a regimen similar to that used in humans, consisting of debulking surgery followed by definitive radiation therapy. Our study found that TuJ‐1 is a useful marker for ONB and that radiation therapy, even in cases of advanced disease, may result in prolonged survival. 相似文献
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以尼克红商品蛋鸡鸡胚和出壳雏鸡为材料，分析了不同出雏时间雏鸡性比例的变化，旨在探讨出雏时间与雏鸡性别的关系及鸡胚在孵化期不同性别生长发育的规律，为精细调控孵化条件和制定鸡苗销售计划提供依据。结果表明，不同时间出壳的雏鸡，性比例存在明显差异(P<0.05或P<0.01),并呈现出明显的规律性。在20d12h之前出壳的母雏比公雏多，20d时母雏比例最高(公母性比值为0.268±0.054)；其后公雏比例增加，到21d时公雏比例最高(公母性比值为2．184±0．090）；此后，母雏比例又逐渐增加。到21d16h时公母比例降低到1.453±0.099;但接近出雏结束时，公雏比例又有所回升，至出雏结束时，公母比例上升为1.75±0.057。 相似文献
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Eckert J 《Schweizer Archiv für Tierheilkunde》2007,149(1):5-14
The history of echinococcosis in Europe includes a period of over 2000 years. Already in antiquity metacestodes (hydatids) of Echinococcus granulosus, the causative agents of Cystic Echinococcosis (CE), were observed in animals and humans. Alveolar Echinococcosis (AE), caused by metacestodes of E. multilocularis, was identified as a disease entity only in the middle of the 19th century. It took about 100 years until it was undoubtedly clarified and accepted that CE and AE are not caused by a single Echinococcus species, but by E. granulosus and E. multilocularis, respectively. In the 20th century significant progress has been achieved in echinococcosis research, including diagnosis, epidemiology, therapy, immunology, molecular biology and other fields. However, CE and AE remain actual problems as in many endemic regions resources and structures are lacking for effective surveillance and control of these zoonoses threatening humans. 相似文献
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本研究鸡传染性法氏囊病冻干卵黄抗体和市售精制液体卵黄抗体分别以10羽份和3 mL超剂量肌肉注射各10只18日龄SPF雏鸡,观察14 d均安全。两种抗体以1羽份剂量分别肌肉注射各20只18日龄SPF雏鸡24 h后,用IBDV强毒TL株攻击,连续观察144 h,2个试验组分别有20/20、17/20的雏鸡健活,剖检没有发现法氏囊病变;二者分别以1羽份剂量连续2 d肌肉注射各20只18日龄已经被IBDV强毒TL株攻击12 h的SPF雏鸡,连续观察96 h,2个试验组有19/20、17/20的雏鸡健活,而预防和治疗试验中的阳性对照组中20只鸡均有法氏囊特征性病变发生（20/20）,且死亡率在85%（17/20）以上,阴性对照组雏鸡均正常。 相似文献