首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献，搜索用时 83 毫秒
1.
抗H9亚型禽流感病毒独特型抗体的制备与应用   总被引：3，自引：0，他引：3  
用纯化的鸡抗H9亚型禽流感病毒（AIV）免疫球蛋白G（IgG）作为抗原免疫家兔，制备兔源多克隆抗独特型抗体（Pab2），同时免疫小鼠，应用细胞融合技术建立了分泌抗AIV独特型单克隆抗体（Mab2）的杂交瘤细胞系。经鉴定，2类抗独特型抗体均能竞争性地抑制鸡抗AIVIgG与AIV的结合。保护试验证实，兔源多克隆抗独特型抗体疫苗较单克隆抗独特型抗体疫苗具有更高的保护作用。  相似文献   

2.
以纯化的禽流感病毒（avian influenza virus,AIV）核蛋白作为捕捉抗原，金标兔抗鸡抗体作为金标二抗，建立检测禽流感血清抗体的胶体金免疫层析试纸条（immunochromatographic strip, ICS）。对标准阳性血清不同滴度进行检测，ICS试验的敏感性高于琼脂扩散试验（AGP）。特异性试验证实，禽流感的ICS试验与新城疫（ND）、传染性支气管炎（IBV）、鸡传染性法氏囊病（IBD）及减蛋综合症（EDS-76）均无交叉反应。同时，用ICS试验和HI试验检测105份鸡血清，两者具有较好的符合性。结果认为：禽流感ICS具有快速简便、特异、灵敏等优点。  相似文献   

3.
为建立通用型禽流感病毒(AIV)快速检测方法,本研究将合成的同义NP (ConsNP)基因克隆于pET30a中,利用大肠杆菌系统进行原核表达,以纯化重组ConsNP蛋白作为包被抗原,建立了检测AIVNP抗体的间接ConsNP-ELISA方法,并优化了反应条件.结果表明:抗原包被量为1μg/孔;封闭剂为5％脱脂乳;一抗血清稀释度为1∶200; HRP标记的兔抗鸡IgG(酶标二抗)稀释度为1∶20 000;阴阳性临界值为:OD49nm值＞0.239.该间接ELISA方法与其他亚型AIV阳性血清均呈阳性反应,与新城疫病毒,马力克病病毒,禽传染性囊病病毒阳性血清无交叉反应,具有良好的特异性和敏感性.本研究为进一步研制通用型的禽流感ConsNP-ELISA检测试剂盒奠定了基础.  相似文献   

4.
通过制备兔抗鸡传染性腔上囊病病毒（IBDV）、小鼠抗IBDV和绵羊抗兔免疫血清,并纯化绵羊抗兔辣根过氧化物酶标记抗体,采用方阵滴定法测定其最佳使用浓度,建立了IBDV双夹心ELISA检测方法。结果显示,包被抗体的最适稀释度为1：160,兔抗IBDV的最佳稀释度为1：200,酶标记抗体的最佳稀释度为1：400。特异性和重复性试验结果表明,该方法特异性强,重复性好,与鸡新城疫、鸡传染性支气管炎和鸡减蛋综合征病毒抗原均不发生交叉反应。对人工感染病例的检测结果表明,建立的双夹心ELISA方法可用于临床快速诊断IBDV。  相似文献   

5.
Dot-ELISA检测H9亚型禽流感病毒的研究   总被引：1，自引：0，他引：1  
本研究采用兔抗H9亚型禽流感病毒（AIV）IgG包被于硝酸纤维素膜,酶标羊抗兔IgG作二抗,建立检测H9亚型AIV的Dot-ELISA法。经方阵试验确定兔抗AIV IgG工作浓度为1∶400,酶标羊抗兔IgG的工作浓度为1∶400。作者建立的Dot-ELISA对AIV的最小检测量为3.35×10-9g。Dot-ELISA与HA和HI、AGP及病毒分离法相比,检测63份临床疑似H9亚型AIV病料,Dot-ELISA检出32份(57.14%),HA和HI检出15份(23.81%),AGP检出11份(17.46%),病毒分离检出38份(60.30%)。用抗H9亚型AIV阳性血清可以阻断Dot-ELISA阳性反应,诊断膜片与鸡新城疫病毒、鸡传染性法氏囊病病毒、鸡传染性支气管炎病毒、产蛋下降综合征病毒不出现阳性反应,证明Dot-ELISA特异性好。分别置室温(25 ℃左右)、4 ℃和-20 ℃下保存1个月后,膜片诊断效果不变,对照反应均成立,该方法重复性好（重复符合率为93.9%）,操作简便（3 h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标准化的新技术之一。  相似文献   

6.
朗德鹅禽流感病毒的分离与鉴定   总被引：2，自引：0，他引：2  
用禽流感病毒ELISA试剂盒对某朗德鹅养殖场的病鹅气管粘液进行了检测，发现5份粘液样本均呈禽流感阳性；随后取相应气管组织材料接种于9～11日龄鸡胚分离病毒．发现尿囊液能使鸡红细胞发生凝集，用禽流感病毒H5、H7、H9标准阳性血清和新城疫病毒、传染性支气管炎病毒、传染性喉气管炎病毒、传染性法氏囊炎病毒抗血清作HI试验，结果禽流感病毒H5亚型抗血清的血凝抑制滴度达到2^7，而禽流感病毒H7、H9亚型及其他病毒抗血清无血凝抑制滴度，说明从朗德鹅分离到的病毒为H5亚型禽流感病毒。  相似文献   

7.
利用纯化的H9N2亚型禽流感尿囊液病毒免疫原免疫Balb/c小鼠,应用杂交瘤技术将免疫鼠脾细胞与小鼠骨髓瘤细胞(SP2/0)融合,用间接ELISA方法检测培养上清是否对H5N1亚型AIV及NDV反应,筛选只针对H9N2亚型AIV的阳性细胞株,经克隆获得1株较高亲和力的杂交瘤细胞株,命名为4E7,用其制备的腹水ELISA效价达到2×105,Western-blotting证明单抗4E7经鉴定为针对75KD血凝素的单抗,HI效价为1:27,且不与新城疫病毒、鸡传染性法氏囊病病毒、传染性脑脊髓炎病毒、鸡传染性支气管炎病毒、H5N1亚型禽流感病毒发生交叉反应,具有较好的特异性。IgG亚型为IgG1。  相似文献   

8.
禽流感病毒单克隆抗体的研制及初步应用   总被引：1，自引：0，他引：1  
接种H5N1亚型禽流感病毒(AIV)鸡胚尿囊液，经甲醛灭活后，差速离心提纯病毒，三次免疫BALB／c小鼠，取免疫小鼠脾细胞与骨髓细胞SP2／0-Ag-14融合。用间接ELISA和血凝抑制(HI)试验筛选阳性杂交瘤细胞株。本研究共获14株能够稳定分泌特异性单克隆抗体(McAbs)的杂交瘤细胞株，其中McAb A6具有HI特性，并可以与重组鸡痘病毒表达的H5亚型AIV HA蛋白反应。用间接EHSA试验测定McAbs与125株AIV分离株的反应性。用反应谱广的McAbs(483和F2)进行交叉反应实验，确定了McAbs可用于夹心ELISA的最佳包被和酶标方式。以McAb483为基础建立的夹心ELISA对175株各亚型的AIV的阳性检出率在90％以上。  相似文献   

9.
检测禽流感病毒抗体的重组核蛋白间接ELISA方法的建立   总被引：9，自引：3，他引：9  
以大肠杆菌系统表达的H9N2亚型禽流感病毒（AIV）核蛋白（NP）为抗原，建立了禽流感间接酶联免疫吸附试验抗体检测技术（NP—ELISA）。对263份待检血清（包括临床收集的243份血清和20份H9N2亚型AIV免疫鸡阳性血清）进行检测，NP—ELISA与琼脂免疫扩散试验（AGP）的总符合率为83．3％，与血凝抑制试验（HI）的总符合率为92％。特异性试验表明，NP—ELISA方法可以检测H5、H7和H9亚型AIV特异性抗体，检测为阳性的血清样品能够被阳性鸡胚尿囊液阻断。敏感性试验证实，NP—ELISA最早可以检测鸡感染后7d的血清样品，并于感染后10d确定100％血清阳性，而AGP检测直到首免后21～28d才出现部分血清阳性，HI检测直到10～14d才出现部分血清阳性，并且NP-ELISA要比HI敏感4～40倍。试验证明，NP—ELISA是检测AIV血清型特异性抗体的一种特异、敏感、快速、经济的血清学检测技术。  相似文献   

10.
H5亚型禽流感病毒间接免疫荧光快速诊断方法的建立   总被引：1，自引：0，他引：1  
本研究以当前严重威胁我国养禽业的高致病性禽流感H5亚型病毒为研究对象，病毒在犬肾细胞（MDCK）上培养增殖，经蔗糖梯度离心对病毒进行纯化，免疫清洁级的新西兰公兔，高免血清经辛酸-硫酸铵法和葡聚糖G50柱纯化，制得第一抗体。以FITC标记的山羊抗兔IgG为第二抗体，通过反应条件的优化，建立了间接免疫荧光快速诊断方法。本法的最佳检测组织为心肌和胰腺，检测时间只需3小时，本法可检出人工感染后36小时尚未表现出临床症状鸡只中的病毒，对禽流感H7亚型、H9亚型病、新城疫、传染性支气管炎和传染性喉气管炎禽出败等病料进行特异性检验结果均为阴性。运用本方法对69个禽场的临床病料进行了检测，检测结果与鸡胚分离法进行比对，9个阳性场（广东省2004年9个原疫点的病料）的9份病料中，检出8份阳性；而鸡胚分离法阴性样品，本法检测结果与之完全相符。本法用于禽流感H5亚型病毒的快速诊断具有快速、简便、敏感、特异、费用低廉和不存在交叉污染等优点，在当前流行的H5亚型高致病性禽流感快速诊断中具有良好的应用前景。  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.
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   

15.
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   

16.
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   

17.
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   

18.
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   

19.
Comparison of different tests to diagnose feline infectious peritonitis   总被引：1，自引：0，他引：1  
  相似文献   

20.
An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号