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1.
猪伪狂犬重组抗原间接ELISA诊断方法的建立   总被引：5，自引：0，他引：5  
本研究应用重组抗原,成功建立了检测猪伪狂犬病病毒血清抗体的间接ELISA诊断方法。通过方阵滴定确定了抗原最适包被量为1.45μg/孔,血清最适稀释度为1∶80,其作用时间为60 min,酶标抗体最适作用时间为60 min。其阳性临界值为OD≥0.123。此外,该抗原不与猪其他疾病的阳性血清反应,具有良好的特异性;批内重复试验,变异系数小于10%。该方法与进口诊断试剂盒的符合率为96.3%。本研究为现地免疫猪群抗体检测和进行PRV流行病学调查提供了一种简便的血清学诊断方法。  相似文献   

2.
间接ELISA检测犬细小病毒病血清抗体方法的建立   总被引：8，自引：0，他引：8  
用犬肾细胞增殖犬细小病毒,经纯化﹑标化作为包被抗原,建立检测犬细小病毒抗体的间接ELISA方法。结果表明:抗原最佳包被浓度5μg/mL;血清最佳稀释度1:40,反应时间45min;酶标抗体最佳稀释度1:2000,反应时间45min;S/P≥0.240判为阳性,≤0.190判为阴性,介于二者之间为可疑。该抗原不与犬瘟热﹑犬传染性肝炎﹑犬冠状病毒病﹑猫泛白细胞减少症病原阳性血清反应;批内重复试验变异系数小于10%,批间重复试验变异系数小于15%;对20份免疫犬血清进行检测,阳性检出率为85%,明显高于血凝抑制试验(65%),符合率为80%。试验结果表明建立的间接ELISA检测犬细小病毒抗体方法特异、灵敏、可重复性好。  相似文献   

3.
为建立鸡毒支原体(MG)种特异性检测的间接ELISA检测方法,本研究以原核表达的PvpA蛋白作为包被抗原,初步建立了检测MG抗体的间接ELISA检测方法.特异性试验结果表明,重组PvpA蛋白与鸡新城疫病毒、鸡传染性支气管炎病毒、滑液支原体、大肠杆菌“O”抗原、禽沙门氏茵“O”抗原阳性血清均不发生交叉反应.该方法的批内变异系数小于8％,批间变异系数小于11％,表明具有较好的重复性.采用该检测方法与进口试剂盒对不同省份送检的鸡血清进行检测比较,两者阳性和阴性符合率均达到90％以上.本研究建立的间接ELISA检测方法为MG抗体检测试剂盒的研制奠定了基础.  相似文献   

4.
采用RT-PCR方法扩增猪瘟病毒(CSFV)E2蛋白主要抗原区基因,克隆入原核表达载体pET32a(+),转化大肠埃希菌BL21构建重组表达菌,经SDS-PAGE和Western blot鉴定目的蛋白得到成功表达。利用纯化的重组蛋白作为包被抗原,通过反应条件优化,建立了间接ELISA抗体检测方法。ELISA抗原最适包被浓度为0.5μg/mL(0.05μg/孔),最佳封闭液为5g/L BSA,待检血清最适稀释度为1∶100,作用时间为1h,酶标抗体最适稀释度为1∶2 000,最适作用时间为45min,室温显色10min。用该方法检测牛病毒性腹泻病毒(BVDV)、猪圆环病毒2型(PCV-2)、猪繁殖与呼吸综合征病毒(PRRSV)、伪狂犬病病毒(PRV)阳性血清结果均为阴性;批内和批间重复试验变异系数分别<5%和<8%,表明本方法具有较好的特异性和重复性。应用△E2-ELISA对350份血清样品进行检测,阳性率为77.71%,高于IDEXX试剂盒检测阳性率(67.14%),与IDEXX试剂盒阳性符合率91.06%,阴性符合率50%,总符合率77.43%。表明本试验建立的间接ELISA方法适于临床CSFV血清抗体的检测。  相似文献   

5.
本研究以纯化的猪繁殖与呼吸综合征病毒(PRRSV)抗原作为诊断抗原,建立了检测PRRSV抗体的间接ELISA诊断方法。该方法检测7种常见猪病病毒(PCV-2、PRV、JEV、PPV、PEDV、TGEV、CSFV)的阳性血清均为阴性;批内重复性试验的变异系数小于5%,批间重复性试验的变异系数小于10%;与商品化ELISA试剂盒相比较,符合率、敏感性和特异性分别为91.1%、90.5%和91.8%。本研究建立的PRRSVELISA检测方法具有良好的重复性、敏感性和特异性。  相似文献   

6.
利用pET32a(+)-Cap重组蛋白作为包被抗原,通过反应条件优化,建立了间接ELISA方法用于检测猪圆环病毒2型(PCV2)抗体。结果表明,抗原最适包被浓度为4μg/mL,最佳封闭液为1%BSA,37℃封闭3 h,血清最适稀释度为1∶200,其作用时间为60 min,酶标抗体最适稀释度为1∶20 000,最适作用时间为30 min,37℃显色15 min,判定血清样品P/N≥2.1,且OD450≥0.228为阳性。该方法与猪瘟、猪口蹄疫、猪脑心肌炎、猪呼吸与繁殖综合征病毒阳性血清反应呈阴性。批内和批间重复性试验结果变异系数均值分别为5.65%和5.81%,表明本方法具有较好的特异性和重复性。应用本方法对123份血清样品进行检测,检测结果阳性率78%,与国产商品化试剂盒检测结果对比,符合率为91.9%,表明本试验建立的间接ELISA方法具有较高的敏感性,适于大规模检测PCV2血清抗体的流行病学调查。  相似文献   

7.
犬细小病毒ELISA抗体检测试剂盒的研制及应用   总被引：1，自引：0，他引：1  
本研究以纯化的犬细小病毒(CPV)重组VP2蛋白为包被抗原,建立了检测CPV抗体的间接ELISA 方法,其抗原包被浓度为0.158μg/mL;待检血清稀释倍数为1:100,作用时间为60 min;羊抗犬酶标抗体稀释倍数为1:2 000,作用时间为60 min;底物作用时间为10 min;判定标准为S/P≥O.282时为阳性,S/P≤0.226时为阴性.该方法与犬瘟热、犬传染性肝炎、犬副粘病毒病、犬波特氏杆茵痛等4种犬常见传染病阳性血清无交叉反应,具有较好的特异性;批内、批间重复试验变异系数均小于15%,显示该方法具有良好的重复性;与商品化CPV ELISA试剂盒进行比较,符合率为96.5%.本研究为现地免疫犬群抗体监测和进行CPV流行病学调查提供了一种简便的血清学诊断方法.  相似文献   

8.
为建立敏感、特异、稳定的传染性法氏囊病病毒(IBDV)VP2结构蛋白抗体ELISA检测方法,本试验以杆状病毒表达系统表达的重组IBDV-VP2结构蛋白为抗原,通过包被VP2单克隆抗体捕捉VP2结构蛋白的形式,建立鸡血清VP2结构蛋白抗体ELISA检测方法(VP2-ELISA)。条件优化结果显示:单克隆抗体最适包被质量浓度为3mg/L,VP2最佳稀释度为1∶20,夹心ELISA效价1∶12.8,被捡血清的稀释度为1∶400。该方法与禽流感、鸡新城疫、鸡马立克氏病、鸡传染性喉气管炎病毒及杆状病毒阳性血清无交叉反应,对琼扩效价为1∶32的IBDV阳性血清的最低检出量为1∶12 800,ROC曲线确定的D450nm临界值为0.236,组内、组间重复性试验变异系数均小于10%。用本试验建立的VP2-ELISA方法与进口IBDV-ELISA抗体试剂盒平行检测100份背景已知的临床鸡血清样品,总符合率VP2-ELISA(97%)高于进口ELISA试剂盒(96%)。本试验为进一步研制鸡传染性法氏囊病病毒VP2结构蛋白抗体ELISA检测试剂盒奠定基础。  相似文献   

9.
为建立蓝舌病病毒(BTV)病原学检测方法,本研究以纯化的BTV-1免疫绵羊和兔子,制备高免血清。以绵羊抗BTV血清为捕捉抗体、以兔抗BTV血清和羊抗兔Ig G-HRP为检测抗体建立了检测BTV的抗原捕获ELISA(AC-ELISA)方法。该方法能够特异性检测BTV,对羊痘病毒、赤羽病病毒无交叉反应,与鹿流行性出血热鸡胚样品有微量交叉反应,但样品OD450nm/阴性对照OD450nm低于阴阳性临界值,不影响结果判定;敏感性试验检测结果表明该方法可以检出102.79 TCID50的病毒;批内和批间重复性试验变异系数分别为1.44%~8.46%和2.26%~12.44%;采用该方法与RT-PCR方法对60份鸡胚样品进行检测,阳性符合率为90%。本研究建立的AC-ELISA方法为BTV抗原检测提供了一种快速、实用的技术手段。  相似文献   

10.
用大肠杆菌表达的牛传染性鼻气管炎病毒（IBRV）重组gD蛋白纯化后作为包被抗原,建立了检测牛传染性鼻气管炎病毒抗体的间接ELISA方法。交叉反应试验表明,该重组抗原与其它常见的5种牛病阳性血清不发生交叉反应;阻断反应试验表明,IBRV病毒悬液能在很大程度上阻断重组抗原与阳性血清的反应,而对阴性血清没有明显影响。在重复性试验中,批内重复的变异系数小于5%,批间重复的变异系数小于15%。与中和试验相比较,符合率、敏感性和特异性分别为84.1%、85.0%和83.4%。应用该诊断方法和本实验室已建立的以IBRV全病毒作为包被抗原的ELISA诊断方法,同时检测了采集于国内11个省份的2012份血清样本,IBRVgD-ELISA检测的平均阳性率为46.0%（926/2012）,而且相对于IBRV全病毒ELISA诊断方法的符合率、敏感性和特异性分别为91.9%、94.2%和90.2%。本研究所建立的IBRVgD-ELISA具有良好的敏感性和特异性,为国内IBR流行病学调查提供了一种快速、简便的血清学诊断方法。  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.
Tropical Animal Health and Production -  相似文献   

14.
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   

15.
Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed.  相似文献   

16.
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   

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 objective of this report was to characterize 20-year changes in proportion of calcium oxalate (CaOx) calculi and struvite calculi in dogs, and associations with breed, age, and sex. In this retrospective study, results of analysis of urinary calculi from dogs were reviewed for specimens received between July 1, 1981, and December 31, 2001. Breed, sex, age, year of submission of the specimen, and mineral type(s) were analyzed statistically. CaOx or Struvite or both were contained in 18,966 of 20,884 (91%) specimens. For both sexes, a 20-year statistically significant increase was observed in the proportion of calculus specimens that contained CaOx. The increase in this proportion was greater in females (1% to 31%) than in males (18% to 82%). From 1998 to 2001, when proportions may have plateaued, the odds of specimens containing CaOx were markedly higher in 18 breeds, markedly lower in 5 breeds, and not significantly different in 13 breeds compared with crossbreds. For both sexes, a 20-year statistically significant decrease was observed in the proportion of calculus specimens that contained struvite. This decrease in proportion was greater for males (79-16%) than for females (97-68%). From 1998 to 2001, when proportions plateaued, the odds of calculi containing struvite were markedly lower in 20 breeds, markedly higher in 1 breed, and not significantly different in 15 breeds when compared with crossbreds. Breed, age, and sex were associated statistically with CaOx or struvite urolithiasis. In conclusion, there appears to have been a long-term increase in the proportion of specimens of canine urinary calculi that contain CaOx as well as a long-term decrease in the proportion of specimens of calculi that contain struvite for both male and female dogs. The rate of change appeared to begin leveling off in the period 1998 to 2001. The recent proportion of dogs with either CaOx- or struvite-associated urolithiasis may depend on breed, age, and sex, and on interactions among these 3 factors.  相似文献   

19.
<正>1.INTRODUCTION Ensuring transportation network security is one Of the most daunting challenges confronting homeland security agencies today.Significant research has been dedicated.To model and analyze the vulnerability of transportation systems,while notably fewer studies propose specific strategies for deploying defensive technologies to safeguard these systems.  相似文献   

20.
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   

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