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1.
氨苄青霉素单抗鉴定与酶联免疫检测方法的初步研究   总被引：6，自引：1，他引：6  
利用碳化二亚胺(EDC)法将氨苄青霉素(AMP)与匙孔嘁血蓝蛋白(m cKLH)偶联制备免疫原免疫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,建立分泌氨苄青霉素单克隆抗体的杂交瘤细胞株。杂交瘤细胞染色体数目为97~104条,3D12株亚型为IgG2 a,用其制备的腹水ELISA效价达到2×106。亲和常数为3×10-10mol/L,抗体相对分子质量为1.54×105,与其他抗生素的交叉反应率小于0.01%。采用间接竞争ELISA方法建立检测AMP的标准曲线,在0.5~100 ng/mL范围内呈线形相关,回归方程为Y=0.056 9X+0.130 8,相关指数R2=0.994 8,以10%抑制率对应的浓度计算,该方法对AMP的检测限为0.3 ng/mL,对市售消毒纯牛奶模拟样品最低检测限为0.4 ng/mL。  相似文献   

2.
采用碳化二亚胺法,将双氯青霉素(dicloxacillin)与牛血清白蛋白偶联制备免疫原,免疫BALB/c小鼠,取免疫小鼠的脾细胞与Sp2/0骨髓瘤细胞融合,间接ELISA筛选,阳性杂交瘤细胞用有限稀释法克隆,获得1株可稳定分泌McAb的杂交瘤细胞5D4-C9-C8;间接ELISA方法测定,其细胞上清抗体效价为1∶300,腹水效价为1∶3.5×105,为IgG1,与其结构类似物邻氯青霉素、苯唑青霉素的交叉反应率分别为17.6%和26.1%,与苄青霉素、羟氨苄青霉素和氨苄青霉素均无交叉反应性。体外传代培养和冻存复苏后抗体分泌稳定。为进一步研制检测双氯青霉素的ELISA试剂盒奠定了基础。  相似文献   

3.
《畜牧与兽医》2014,(12):15-18
用酸酐法将氨苄青霉素(Ampicillin,AMP)偶联于载体蛋白牛血清白蛋白(BSA)和匙孔血蓝蛋白(Keyhole Limpet Hemocyanin,KLH),合成了免疫抗原AMP-KLH和检测抗原AMP-BSA,并进行了鉴定;用免疫抗原AMP-KLH免疫BALB/c小鼠,应用杂交瘤技术筛选出了3株分泌抗AMP单克隆抗体的杂交瘤细胞株。特性分析显示,单克隆抗体2C3、4E5和4C7的腹水效价分别为1∶1.02×106、1∶5.12×105和1∶5.12×105,其中2C3的AMP半数抑制浓度(IC50)为9.3 ng/m L,与青霉素类抗生素有明显交叉反应,其他类别抗生素的交叉反应率均小于0.01%。  相似文献   

4.
应用重氮化方法制备了磺胺氯吡嗪钠(sulfachloropyrazine sodium,SPZ)完全抗原SPZ-OVA,将成功偶联的人工全抗原(SPZ-OVA)免疫BALB/c小鼠,免疫4次后,经间接ELISA检测,1号小鼠血清抗体效价达到1:6.4×104。取该小鼠脾细胞与SP2/0细胞融合。利用ELISA方法对融合后的杂交瘤细胞上清进行检测,筛选出能够稳定分泌抗体的细胞株:D9。将细胞株D9扩大培养后,经腹腔注射小鼠使其产生腹水。腹水单抗经纯化后,通过间接竞争ELISA对该单克隆抗体的效价、半数抑制浓度以及特异性进行检测。结果表明,所制备的单克隆抗体效价为1:4×105,半数抑制浓度为326 ng/m L,特异性良好。  相似文献   

5.
主编 《中国兽药杂志》2010,44(10):29-33
采用碳二亚胺(EDC)法将氟罗沙星(FLE)与载体蛋白BSA、OVA分别偶联合成人工抗原BSA-FLE、OVA-FLE,通过聚丙烯酰胺凝胶电泳法和紫外分析法对合成的人工抗原进行了鉴定。BSA-FLE用作免疫抗原免疫Balb/C小鼠,OVA-FLE用作检测抗原包被酶标板检测抗体效价。免疫小鼠的脾细胞与SP2/0细胞融合获得了5株(4F11、2C3、8G2、3C1、2G8)稳定分泌抗FLE抗体的杂交瘤细胞株,抗体亚型鉴定均为IgG1。将3C1接种小鼠制备腹水,用辛酸-硫酸铵法对腹水进行纯化,纯化后腹水效价1∶1 024 000。抗体分子量160.766 KD,亲和常数为8.57×108M-1。  相似文献   

6.
用小鼠骨髓瘤细胞(SP2/0)与经篮舌病病毒免疫的BALB/C小鼠脾细胞进行融合,产生杂交瘤细胞。以免疫荧光(间接法)和ELISA(间接法)进行杂交瘤抗体分泌检测,用有限稀释法进行3次克隆,获得5株抗体分泌稳定、产量高的单克隆抗体杂交瘤细胞株。  相似文献   

7.
采用混合酸酐法将短裸甲藻毒素(brevetoxins,BTX)分别与卵清蛋白(OVA)和牛血清蛋白(BSA)偶联制备人工抗原。以BTX-BSA作为免疫抗原免疫BALB/c小鼠,通过细胞融合-ELISA筛选抗短裸甲藻毒素的单克隆抗体,并对其特异性和灵敏性进行了鉴定。结果获得了1株可分泌短裸甲藻毒素单克隆抗体的杂交瘤细胞株3B4,该株杂交瘤细胞腹水抗体效价达10-5,与其他类似物没有交叉反应。该抗体为建立短裸甲藻毒素免疫学检测方法奠定了基础。  相似文献   

8.
制备抗促乳素(prolactin,PRL)的单克隆抗体。应用PRL纯品免疫BALB/c小鼠,被免疫小鼠的脾细胞与SP2/0骨髓瘤细胞融合,间接ELISA筛选阳性克隆,将阳性克隆多次有限稀释得到抗PRL阳性单克隆细胞株,杂交瘤细胞上清分离法和诱生腹水法制备单克隆抗体,测其效价。共建立5株持续分泌抗体的杂交瘤细胞,分别命名为AA5B1、AB6C8、BB3C1、CA6E4、CE2H6。杂交瘤细胞株冻存4个月后复苏培养,形态良好,生长旺盛。培养液上清及腹水效价的OD值略有升高,说明杂交瘤细胞株稳定。5株杂交瘤细胞染色体众数均为99～105,并可见到中部和亚中部着丝点的骨髓瘤细胞标志染色体,说明5株杂交瘤细胞确为SP2/0骨髓瘤细胞与被免疫小鼠的脾细胞融合而产生的。  相似文献   

9.
己烯雌酚完全抗原及其单抗的制备与分析   总被引：1，自引：0，他引：1  
为建立己烯雌酚免疫学检测技术,试验采用混合酸酐法制备完全抗原DES-HS-BSA,并对制备产物进行紫外光谱、红外光谱、SDS-PAGE、MS分析鉴定,以DES-HS-BSA免疫Balb/c小鼠,用ELISA试验测定抗体.结果表明:半抗原衍生物DES-HS已被成功地偶联到牛血清白蛋白上,计算得每分子牛血清白蛋白上结合26个DES-HS;ELISA试验结果证实免疫小鼠血清中含有抗己烯雌酚的抗体,将免疫好的小鼠脾脏细胞和SP2/0细胞融合,筛选出了1株能稳定分泌己烯雌酚单抗的杂交瘤细胞株18G5,抗体亚型为IgG3.  相似文献   

10.
琥珀酸酐法将雌二醇(E2)衍生化,再用碳二亚胺法(EDC)将半抗原与BSA和OVA偶联,制备免疫原和检测抗原,紫外扫描和红外光谱鉴定表明人工抗原合成成功,E2与BSA偶联比为12.3∶1。将E2-BSA免疫BALB/c小鼠,细胞融合技术筛选抗E2杂交瘤细胞株,制备单克隆抗体。结果表明,筛选的3株杂交瘤细胞E2D5、E3F7和E4A6抗体效价均在8×104以上。以E3F7细胞株建立间接竞争ELISA(icELISA)标准曲线,其线性范围为0.0666.2μg/L,检测限和IC50值分别为0.03和0.76μg/L,除与去氢甲睾酮22.9%的交叉反应率外,与其他化合物无交叉反应。本研究为开发ELISA试剂盒,检测食源性动物产品中E2残留奠定基础。  相似文献   

11.
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   

12.
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   

13.
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   

14.
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   

15.
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   

16.
Comparison of different tests to diagnose feline infectious peritonitis   总被引：1，自引：0，他引：1  
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17.
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   

18.
Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein  相似文献   

19.
正We are pleased to publish the second special issue on avian brood parasitism and to be responsible guest editors for the two special issues of Chinese Birds (Vol. 3, No. 4, 2012 and Vol. 4, No. 1, 2013), entitled "Avian Brood Parasitism - A Growing Research Area in Behavioral Ecology". The first issue was published in December 2012. The goal of the two special issues is to publish accumulated knowledge and some of the recent developments in  相似文献   

20.
Tropical Animal Health and Production -  相似文献   

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