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1.
双抗体夹心ELISA检测牛病毒性腹泻病毒的研究   总被引:21,自引:3,他引:18  
建立了从粪便中检测牛病毒性腹泻(BVD)病毒抗原的双抗体夹心ELISA。试验方法的最佳工作条件为,抗体包被量为100μg/孔,酶标抗体的浓度为1:400。对不同地区牛、羊、鹿粪样检测结果,牛的BVDV感染率为16.5% ̄89.0%,羊为14.6% ̄83.3%,鹿为19.6% ̄44.4%,并且从许多地区的牛、羊同时检测到BVDV,表明本病在国内某些地区存在着严重的感染。  相似文献   

2.
为建立检测牛病毒性腹泻病毒抗体的间接ELISA检测方法,利用大肠杆菌表达系统成功表达E2蛋白并纯化,利用纯化后的蛋白作为包被抗原,用方阵法对影响ELISA试验的各个因素进行优化,并进行特异性、敏感性和重复性试验。确定E2抗原最佳包被浓度为1μg/mL,最佳血清稀释度为1∶40,最佳封闭液为1%明胶,最佳血清作用方式为37℃作用30 min,酶标抗体的最佳作用方式为1∶2000稀释、37℃作用30 min;最佳底物作用时间为室温20 min,阳性临界值为OD450nm≥0.255。与血清中和试验和商品化试剂盒进行比较,总符合率分别为98%和96%,批内和批间重复性试验的变异系数均小于10%。该方法与其它常见牛病毒阳性血清均无交叉反应。本研究成功建立了间接ELISA方法,为进一步用于大批量牛血清样本的抗体检测和流行病学研究奠定了基础。  相似文献   

3.
为了调查某规模化奶牛场犊牛腹泻的发病原因,试验采用血清抗体检测、细菌分离培养、寄生虫卵镜检和病毒核酸检测方法对腹泻犊牛的血样和粪便进行了检查。结果表明,该奶牛场牛病毒性腹泻/黏膜病病毒抗体阳性率为65.00%,牛轮状病毒抗体阳性率25.00%,牛冠状病毒抗体阳性率15.00%;分离到1 株结肠弯曲杆菌和1 株空肠弯曲杆菌;所检腹泻犊牛粪便和血液样品中,4 份粪便样品和1 份血液样品检出牛冠状病毒,2 份粪便样品检出轮状病毒,1 份粪便样品和1 份血液样品检出牛病毒性腹泻/黏膜病病毒,同一份粪便样品中同时检出牛病毒性腹泻/黏膜病病毒和牛冠状病毒;通过镜检,球虫卵囊检出率77.78%,在1 份粪样中发现蛔虫卵,2 份粪样中发现其他线虫卵。该牛场流行性腹泻的原因可能为夏季高温潮湿引起犊牛免疫力下降导致的多病原体感染。  相似文献   

4.
检测猪瘟病毒(HCV)抗体的实验室诊断方法有琼脂扩散试验、对流免疫电泳试验、补体结合试验、间接酶标抗体试验和间接荧光抗体试验等。但这些实验的敏感性都比较低,而且不能区分出HCV和牛病毒性腹泻病毒(BVDV)的抗体。据报道,用酶标抗体和荧光抗体血清中和试验检测HCV  相似文献   

5.
采用sf9昆虫细胞-杆状病毒系统重组表达牛病毒性腹泻病毒P80蛋白抗原,经纯化鉴定后将P80蛋白用胶体金标记作为示踪抗原,未标记的P80抗原作为捕获抗原,并以羊抗牛IgG抗体作为质控抗体,建立了牛病毒性腹泻病毒抗体快速诊断试纸条。试验结果表明,该试纸条检测灵敏度高,特异性良好,与牛口蹄疫病毒、牛传染性鼻气管炎病毒无交叉反应。使用建立的胶体金试纸条和IDEXX牛病毒性腹泻病毒ELISA抗体检测试剂盒同时检测80头份牛血清,阳性结果符合率为86.7%,阴性结果符合率为100%,总符合率为95%。该试纸条可用于临床牛病毒性腹泻病毒抗体的诊断和检测。  相似文献   

6.
以牛支原体(Mycoplasma bovis)全菌蛋白经超声波裂解后的裂解物作为包被用抗原,建立检测牛支原体血清抗体的间接ELISA方法。通过棋盘滴定法确定抗原最适包被浓度、待检血清最佳稀释度,同时对抗原的包被方式、封闭剂和封闭时间、酶标二抗最佳工作浓度、一抗和二抗最佳作用时间、血清稀释液、底物显色时间进行优化。用该方法测定10份阴性血清的OD450值计算出阴性临界值,并对牛布氏杆菌病、牛病毒性腹泻黏膜病抗血清进行检测。结果:抗原最适包被浓度为200μg/mL,待检血清最佳稀释度为1∶100,阴性临界值为0.327。用该方法检测牛布氏杆菌病、牛病毒性腹泻黏膜病抗血清均无交叉反应,表明该间接ELISA具有很好的特异性。  相似文献   

7.
用纯化牛病毒性腹泻病毒免疫蛋鸡制备出的卵黄抗体作为包被抗体,采用自制的单抗为一抗,建立牛病毒性腹泻病毒抗原捕获ELISA方法。通过试验确定,抗牛病毒性腹泻病毒卵黄抗体最佳包被浓度为1:50;McAb最适稀释浓度为1:10,HRP-羊抗鼠IgG工作浓度为1:800。通过引入牛病毒性腹泻病毒质控血清进行质控检验,该方法所得检测结果均在质量控制范围内,达到预定标准化要求。标准化的抗原捕获ELISA方法具有特异、灵敏、可靠、方便、快捷等特点,可广泛应用推广,为我国牛病毒性腹泻病毒监测提供了行之有效的技术手段。  相似文献   

8.
由于牛病毒性腹泻病毒与猪瘟病毒之间存在共同抗原和交叉免疫,选择A、B两组各15头牛病毒性腹泻病毒检测为阴性的牦牛,分别注射牛病毒性腹泻疫苗和猪瘟疫苗,在不同时间段(第7、14、30、60、90、180天)采集两组牦牛血清,应用酶联免疫吸附试验检测抗体,结果显示两种疫苗都可以使牦牛机体产生抗体,并且牛病毒性腹泻病毒疫苗比猪瘟疫苗整体抗体水平高,  相似文献   

9.
用牛病毒性腹泻病毒(BVDV)单克隆抗体包被酶标板,以兔多克隆抗体作为夹心抗体,建立BVDV抗原捕获ELISA(AC-ELISA)检测方法,优化反应条件并对该方法的稳定性等指标进行了测试和评价。结果表明,单抗最佳包被质量浓度为5μg/mL,兔多抗血清最佳质量浓度为10μg/mL;单抗在4℃包被12~24 h,多抗在37℃作用1 h为双抗体的最佳反应条件;酶标抗体最适稀释度1∶10000,最适作用时间为1 h;采用1%BSA和1%明胶分别在抗体包被后和加入待检抗原反应后进行两次封闭效果好。用AC-ELISA方法检测临床采集的11份牛腹泻病料和12份健康牛组织样品,同时以病毒分离和RT-PCR检测方法做对比,3种方法符合率很高。研究表明AC-ELISA方法稳定性好,可用于BVDV的临床快速检测。  相似文献   

10.
为了解河北省及周边地区奶牛场牛病毒性腹泻流行情况,从河北、天津、北京共8个地区奶牛场采集妊娠流产奶牛血清样品164份、腹泻犊牛粪样79份,采用ELISA法检测血清样品牛病毒性腹泻病毒(BVDV)抗体,PCR方法检测粪样BVDV抗原E2基因。结果表明,不同奶牛场妊娠流产奶牛均可感染BVDV,血清抗体最低阳性检出率为4.0%,最高阳性检出率为100%,平均检出率为46.95%。腹泻犊牛粪样BVDV抗原E2基因最低阳性检出率为10%,最高阳性检出率为100%,平均检出率为40.5%。说明河北省及周边地区妊娠流产奶牛与腹泻犊牛群中均存在BVDV感染。  相似文献   

11.
试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。  相似文献   

12.
Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.  相似文献   

13.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

14.
Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.  相似文献   

15.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

16.
The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.  相似文献   

17.
Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.  相似文献   

18.
The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.  相似文献   

19.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

20.
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