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1.
利用大肠杆菌表达的牛病毒性腹泻病毒(BVDV)重组E2蛋白作为抗原,建立了检测BVDV血清抗体的间接Dot-ELISA.最佳工作条件为:E2蛋白抗原的最适包被质量浓度2.0 mg/L(2.0 ng/点),酶标抗体的工作浓度为1:500,血清稀释度为1:100,3%明胶-TBS作为封闭液,封闭45 min效果最佳.通过重复性试验、交叉试验、特异性试验和稳定性试验等证明,该方法重复性好、特异性强、灵敏度高;与用BVDV全病毒为抗原的IDEXX ELISA试剂盒相比,特异性96.67%,灵敏度90%,符合率95%.应用建立的检测方法对河北省4个奶牛场采集的178份腹泻奶牛血清样本进行检测,结果BVDV血清抗体阳性率40.45%,与IDEXX ELISA试剂盒的检出率无明显差异.  相似文献   

2.
从持续感染牛白血病病毒（BLV）的羊胎肾细胞（FLK-BLV）中提取前病毒DNA,用PCR方法扩增编码牛白血病病毒gp51蛋白的env（gp51）基因,序列测定结果表明扩增片段全长828 bp。将扩增基因插入原核表达载体pET-32a构建pET-32a-gp51重组质粒,转化BL21（DE3）进行诱导表达,SDS-PAGE电泳结果表明重组融合蛋白大小约为43 000,Western-blot结果表明重组表达蛋白具有免疫反应性。以纯化的重组蛋白gp51作为抗原,建立检测BLV抗体的间接ELISA诊断方法。结果表明,抗原gp51的最佳包被浓度为2.19μg.mL^-1,血清的最佳稀释倍数为1∶80,阳性判定标准确定为样品OD450 nm大于0.451。用该方法对12份参考血清进行检测,准确率为91.67%。  相似文献   

3.
为评价牛γ-干扰素ELISA检测方法检测牛结核的效果及国产试剂盒的检测效果,本试验首先将国产试剂盒与Prionics试剂盒对42份相对阳性的样品和105份相对阴性的样品进行对比研究。然后对5个规模化牛场的3000头奶牛首先进行国产单纯结核菌素颈部皮内变态反应试验,3天后选取皮内变态反应阳性和可疑及部分阴性牛共418头,进行牛γ-干扰素试验。结果国产试剂盒对阳性和阴性样品的检测敏感性和特异性分别为95.2%和100%,与Priobics试剂盒的符合率为99.3%。表明国产试剂盒与进口试剂盒的检测能力一致,牛γ-干扰素检测方法准确可靠。5个牛场的3000头奶牛单纯结核菌素颈部皮内变态反应试验阳性为138头,可疑105头。γ-干扰素试验对418头奶牛的检测,其中阳性74头,与颈部皮内变态反应（可疑牛暂时视为阴性）的符合率为60.5%。  相似文献   

4.
小反刍兽疫YN PPRV C-ELISA抗体检测试剂盒由云南省出入境检验检疫技术中心自主研发,采用这一试剂盒与法国IDVET研制的小反刍兽疫病毒C-ELISA抗体检测试剂盒,分别对田间牛羊的661份血清样本进行对比检测试验,并使用YNPPRV C-ELISA抗体检测试剂盒对15份检出的已知背景血清进行批内和批间可重复性试验。结果显示自行研制试剂盒相对国际通用试剂盒ID VET PPRV C-ELISA的特异性、敏感性、符合率分别达到97.96%、97.82%和98.34%,两者一致性用Kappa值表示,Kappa=0.97。批内试验强阳性、弱阳性变异系数为0%～0.05%。结果表明,YN PPRV C-ELISA抗体检测试剂盒的敏感性、特异性、批内和批间重复性均较高,具有高度的可靠性,在小反刍兽疫的诊断检测中具有良好的实用价值。  相似文献   

5.
为比较两种商品化赤羽病抗体ELISA试剂盒的敏感性和特异性,以及ELISA和微量血清中和试验(SNT)的符合率,采用两种ELISA试剂盒和血清中和试验,对690份澳大利亚纯种荷斯坦奶牛血清进行牛赤羽病检测。试验结果显示:法国某公司生产的赤羽病ELISA试剂盒敏感性为93.42%,特异性为81.23%,与SNT间的Kappa值为0.615,为高度符合;日本某公司生产的赤羽病ELISA试剂盒敏感性为39.47%,特异性为96.1%,与SNT间的Kappa值为0.427,为中度符合。比较结果表明:日本某公司生产的试剂盒敏感性较低,容易漏检,不适合用于牛赤羽病初筛;在出入境检疫中,可以采用高通量、半自动化、敏感性高的ELISA方法,对血清样本进行筛选,再采用特异性强的SNT试验进行辅助验证。  相似文献   

6.
《畜牧与兽医》2016,(2):96-98
为了验证云南出入境检验检疫局检验检疫技术中心自主研发的蓝舌病病毒C-ELISA抗体检测试剂盒(YN BTV C-ELISA)的可应用性,将YN BTV C-ELISA抗体检测试剂盒与美国IDEXX公司研制的蓝舌病病毒VP7蛋白抗体ELISA检测试剂盒对田间牛羊的1 028份血清样本进行对比检测试验,并使用YN BTV C-ELISA抗体检测试剂盒对15份检出的已知背景血清进行批内和批间可重复性试验。YN BTV C-ELISA抗体检测试剂盒相对IDEXX公司蓝舌病病毒ELISA抗体试剂盒的符合率、敏感性、特异性分别是98.44%、98.03%、98.85%,两者一致性用Kappa值表示,Kappa=0.97。批内试验强阳性、弱阳性变异系数在0.23~1.81之间,阴性血清变异系数在3.94~6.64之间,批间试验强阳性、弱阳性变异系数在0.26~2.35之间,阴性血清变异系数在3.55~8.51之间。结果表明:YN BTV C-ELISA抗体检测试剂盒的符合率、敏感性、特异性均较高,具有高度的可靠性、一致性和真实性。YN BTV C-ELISA抗体检测试剂盒的批内和批间有高度的重复性,在蓝舌病的诊断中具有良好的实用价值。  相似文献   

7.
以基因工程表达的非洲猪瘟病毒VP73蛋白作为包被抗原,建立了间接ELISA方法,用以检测猪血清中抗非洲猪瘟VP73蛋白的抗体。该方法对非洲猪瘟标准阳性血清的检测灵敏度可以达到1∶2 560,与同类进口ELISA试剂盒相当。此方法只特异性检出非洲猪瘟阳性血清,而对猪传染性胸膜肺炎等5种猪传染病阳性血清的检测结果均为阴性,表明其具有良好的特异性。批内和批间重复性试验结果发现,检测同一份血清的变异系数小于10%,表明其重复性较好。包被好的酶标板37℃放置5d后,对同一份血清的检测敏感性无明显变化,初步表明其稳定性较好。利用建立的间接ELISA方法和进口ELISA试剂盒分别对150份血清样品进行非洲猪瘟血清抗体检测,结果表明本方法的特异性和敏感性分别为99.1%和94.3%,2种方法检测结果的符合率为98%。以上试验表明,本试验建立的间接ELISA方法具有良好的特异性和敏感性、较好的重复性和稳定性,可以满足临床检测的需求。  相似文献   

8.
孕酮ELISA试剂盒在牛奶乳汁检测中的初步应用   总被引：1，自引：0，他引：1  
本实验利用自建的奶牛乳汁孕酮竞争ELISA法(酶标抗原法)研发了奶牛乳汁孕酮ELISA试剂盒。通过对试剂盒的特异性、灵敏性、重复性、稳定性和回收率等主要性能进行优化试验。结果表明:试剂盒可以在4℃条件下保存9个月,与睾酮、雌三醇、雌二醇无特异性反应,灵敏度为0.14 ng/mL,批内差异为4.11%,批间差异为6.26%,回收率为93.80%~107.60%,变异系数为5.65%。应用该试剂盒检测乳汁孕酮诊断奶牛妊娠与未妊娠的准确率分别为93.3%和95.8%。因此,该试剂盒性能良好,妊娠诊断效果理想,为今后在奶牛生产中的广泛应用奠定基础。  相似文献   

9.
采集新疆地区牛血清90份、羊血清76份,以英国参考实验室APHA竞争性ELISA试剂盒检测结果为确诊标准。应用A、B、C、D 4个不同国产商品化布鲁菌cELISA抗体检测试剂盒对上述样品进行对比检测。结果表明4个试剂盒检测同一批样品结果存有差异,同一厂家试剂盒检测牛、羊血清结果也有差异。C试剂盒无论检测牛或羊血清特异性都最高,敏感性也较好;A试剂盒检测牛或羊血清特异性都最低。4个试剂盒与英国参考实验室cELISA抗体检测试剂盒符合度均较好,敏感性均很高,但特异性参差不齐,最高达100%,最低仅60.53%。结果提示,要重视检测试剂盒特异性指标,降低布病的误判率,减少不必要的经济损失。  相似文献   

10.
应用自制牛结核病PCR诊断试剂盒对贵州省某奶牛场共计150份血样、奶样标本中结核分枝杆菌进行检测,结果显示:PCR试剂盒对111份奶样标本进行检测,8份为阳性,阳性检出率7.2%;PCR试剂盒对39份血样标本的检测,12份为阳性,阳性检出率为30.76%。本试验共计检测贵阳附近某奶牛场1 200头份奶牛,先采用PPD检测牛结核病阳性牛和可疑牛,再用PCR法进行检测,PCR检出阳性牛共计20头份,阳性检出率为1.67%;PPD检出阳性牛24头份,检出率为2%。总之,PCR试剂盒在检测牛结核病不同标本中显示出快速、特异等优点,这为今后牛结核病的检测工作提供了一条新途径。  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.
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   

17.
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   

18.
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   

19.
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   

20.
<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   

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