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1.
用赭曲霉素A(OA)-BSA合成抗原免疫Balb/c小鼠,取免疫鼠脾细胞与sp2/0细胞融合,通过克隆和ELISA法筛选,建立三株泌抗OA单克隆抗体杂交瘤细胞株(3C12,1D12和2G7)。用间接ELISA法测定,细胞上清液抗体效价为128^×(3C12)和64^×(1D12和2G7)腹水抗体效价为10^-7(3C12,1D12)和10^-6(2G7)。三株单抗(McAb)均属IgG类,分泌抗体  相似文献   

2.
具有中和活性的减蛋综合征病毒单抗的研制   总被引:1,自引:0,他引:1  
用减蛋综合征病毒毒株WPDV205纯化抗原免疫BALB/c小鼠,在最后一次免疫后第3天取脾细胞与SP2/0细胞在PEG-4000的作用下融合。用间接ELISA初步筛选出阳性克隆10株(4B1、1D4、2D6、3D6、3A5、2A1、3C6、3C1、1B3和2C5)。这些杂交瘤细胞经克隆化和再次检测后生产腹水,用微量细胞中和试验筛选出具有中和活性的单克抗隆体4株(4B1、3C1、3C6和2C5),其  相似文献   

3.
抗鸡传染性法氏囊病病毒单克隆抗体中和株的建立   总被引:6,自引:1,他引:5  
将纯化的鸡传染性法氏囊病病毒(IBDV)组织毒免疫的BALB/c鼠脾细胞与SP2/0骨髓瘤细胞融合,用ELISA,病毒中和试验检测分泌抗体,获得了6株稳定分泌抗IBDV单隆抗体的杂交瘤细胞,命名为NIB-1,NIB-2,NIB-3,NIB-4,NIB-5,NIB-6;6株MCAb均具ELISA特性和免疫沉淀特性,亚类鉴定表明,前4株属于IgG2a,后2株属于IgG1。杂交瘤细胞冻存6个月后复苏,均  相似文献   

4.
抗绵羊肺炎支原体单克隆抗体杂交瘤细胞株的建立   总被引:1,自引:0,他引:1  
用绵羊肺炎支原体抗原免疫的BALB/C小鼠,取血清抗体阳性免疫鼠的脾细胞与SP2/O细胞在聚乙二醇作用下进行融合,产生杂交瘤细胞、用间接ELISA法对杂交瘤一长孔上清液进行检测,筛阳性交瘤细胞株。  相似文献   

5.
用绵羊肺炎支原体(MO)抗原免疫的BALB/C小鼠,取血清抗体阳性免疫鼠的脾细胞与SP2/O细胞在聚乙二醇作用下进行融合,产生杂交瘤细胞、用间接ELISA法对杂交瘤细胞生长孔上清液进行检测,筛选阳性杂交瘤细胞株.以有限稀释法克隆1~2次,得到6株分泌抗绵羊肺炎支原体抗原的单克隆抗体(McAb)杂交瘤细胞株.选择抗体分泌较高的14A6、7A27、B38进行了较详细的研究,结果表明,其核内染色体数为94~96.抗体属性是:1株为IgG2a、2株为IgG1.用间接ELISA法对3株McAb作符异性分析、该McAb只特异地与绵羊肺炎支原体抗原发生反应,而不与猪肺炎支原体、山羊无乳症支原体抗原发生反应.将杂交瘤细胞注射BALB/C小鼠诱生腹水.其抗体效价提高100倍.杂交瘤细胞在液氮中保存1~2年后复苏仍能稳定地分泌抗绵羊肺炎支原体McAb。  相似文献   

6.
抗兔轮状病毒单克隆抗体杂交瘤细胞株的建立   总被引:1,自引:0,他引:1  
用小鼠骨髓瘤细胞与经兔轮状病毒免疫的BALB/c系小鼠脾细胞进行融合,融合成功率为22%。经间接酶联免疫吸附试验和斑点酶联免疫筛选杂交瘤细胞,阳性率为29%左右。对其中6株强阳性的杂交瘤细胞进行了亚克隆,获得高产亚克隆杂交瘤细胞,用BALB/c小鼠生产了腹水,鉴定后选取2株强阳性高效价的杂交瘤细胞,以ELISA阻断试验检验其分泌抗体的特异性,以直接ELISA测其敏感性,用琼脂双扩散试验鉴定类和亚类  相似文献   

7.
一种快速有效制备单克隆抗体的方法   总被引:3,自引:0,他引:3  
以人绒毛膜促性腺激素(hCG)为抗原,对BALB/c小鼠一次脾内直接注射免疫,免疫后6d,取其血清进行抗体检测,结果12800倍稀释度仍呈阳性。取此免疫小鼠脾细胞与同系小鼠骨髓瘤细胞NS-1进行细胞融合。融合后,将经检测抗体阳性的细胞孔置于显微镜下,挑选单个克隆进行克隆化,得到2株单克隆抗体细胞株2A8、4E6。将其分别接种于BALB/c小鼠腹腔中,收获的腹水效价分别达10-6和10-5。  相似文献   

8.
利用猪繁殖与呼吸道综合征病毒(PRRSV)国内分离株J1,采用反复差速离心法制备免疫抗原,长程免疫法免疫BALB/c小鼠,用间接ELISA方法检测抗体,通过细胞融合技术,并经3次亚克隆获得了10株能稳定分泌抗PRRSV单抗的杂交瘤细胞单克隆株(A1D7H10,A1D7H11,A1E7H9,A1E7D9,A2D8E7,A2D8B11,B3D11D6,B2G9A9,B2G9F2)。这些细胞经体外连续传  相似文献   

9.
用从健康猪胸腺提取的 T 淋巴细胞作抗原免疫 B A L B/c 小鼠。取免疫鼠脾细胞与 S P2/0 骨髓瘤细胞融合,经 3 次克隆,共获得 10 株稳定分泌抗体的杂交瘤细胞。以细胞反应特异性试验、 W esternblotting 分析、流式细胞计数( F C M )等方法,确定了对猪 T 细胞表面抗原特异性反应的单抗( M c Abs),其中 2 D9 、3 B1 0 可以识别相对分子质量 50 000 的 T 细胞膜蛋白,能标记 96% 的胸腺细胞、89% 的外周血 T 淋巴细胞、87% 的脾 T 淋巴细胞,其特性与猪 C D2 抗原相符;另一株 1 G1 2 可识别相对分子质量 63 000 的 T 细胞膜蛋白,能标记 98% 的胸腺细胞、91% 的外周血 T 淋巴细胞、78% 的脾 T 淋巴细胞,其特性与猪 C D5 抗原相符。  相似文献   

10.
以纯化的兔出血症病毒免疫BALB/C鼠,应用杂交瘤技术获得了分泌抗RHDV单抗杂交瘤细胞17株,其中有3株杂交瘤细胞分泌单抗的ELISA效价为1:409600,1:4096;5株单抗具有血凝抑制活性。兔抗RHDV高兔血清对17株McAb的ELISA抑株为IgG2b。各株McAb与欧洲棕色兔病病毒无抗原交叉反应,也无沉淀反应特性。  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.
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.
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   

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.
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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