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 62 毫秒
1.
应用单抗竞争ELISA快速检测肠炎沙门氏菌   总被引:1,自引:0,他引:1  
本研究应用抗肠炎沙门氏菌 O9抗原的酶标单抗 3- 4 7- 2 6和肠炎沙门氏菌脂多糖( LPS)建立了快速检测肠炎沙门氏菌的竞争ELISA方法 ,该法通过样品中的肠炎沙门氏菌 L PS与微量板包被的肠炎沙门氏菌 LPS竞争结合酶标抗体 ,以检测样品中的肠炎沙门氏菌。通过与国标法对样品的比较检测结果表明 ,该法可检测出每克粪便中 5×1 0 4 CFU的肠炎沙门氏菌 ,国标法可检出 4.9× 1 0 1 CFU/g粪便。竞争 EL ISA( C- ELISA)方法的敏感性和特异性分别为 94.4%和 97.7%。并进一步讨论了该方法建立时的选择条件和应用意义  相似文献   

2.
检测肠炎沙门氏菌ELISA方法的建立与应用研究   总被引:10,自引:0,他引:10  
从已建立的抗沙门氏茵单抗中筛选出适用于肠炎沙门氏菌ELISA检验的3—47—26单抗.采用筛选强阳性杂交瘤细胞株制备了高效价的3—47—26单抗.改进用辣根过氧化物酶标记高效价单抗的方法,进行了HRP标记单抗的免疫生物学特性的鉴定。确定了包被浓度和酶标抗体的工作浓度:HRP-3—47—26为1:100;LPS的包被浓度为400ng/mL。试验中采用LPS与多聚赖氨酸的结合,解决了包被过程中的解吸附作用,增强了包被的稳定性,同时也减少了假阳性反应,优化了包被过程。建立了检测肠炎沙门氏菌的抗原竞争ELISA方法,利用样品中的LPS抑制酶标抗体与包被的LPS结合,以降低底物反应的颜色从而达到检测的目的。通过对210份肠炎沙门氏菌感染鸡泄殖腔棉拭子、羽毛和组织样品先筛选后鉴定的方法进行检测具有明显的抑制作用,通过与国标法对大量的样品检测结果比较表明,竞争ELISA方法的检出率为18.09%;检出阳性样品36份,国标法检出率为17.14%;两者符合率为97.14%。竞争ELISA敏感性和特异性分别为94.4%和97.7‰从而为肠炎沙门氏菌的检测提供了一种敏感、特异的检测方法。  相似文献   

3.
研制出两株能与鸡组织、鸡蛋和环境样品中肠炎沙门氏菌反应的单克隆抗体(以下简称单抗),并以此建立了能特异检测肠炎门沙氏菌的快速酶联免疫吸附试验和凝集试验。ELISA和常规微生物学分离定系统能检出100%鸡蛋和99.8%不同野外和人工接种样品中的炎沙门氏菌,滴定试验表明ELISA可检出10^4个细菌。  相似文献   

4.
氯霉素间接竞争ELISA(ciELISA)检测方法的建立   总被引:31,自引:4,他引:27  
以人工合成的氯霉素-牛血清白蛋白(CAP-BSA)为包被抗原,氯霉素(CAP)为竞争半抗原,两者与一定量的抗CAP-McAb反应。结果表明,理想的包被抗原质量浓度为1.25mg/L,抗CAP-McAb稀释倍数为1∶12000,酶标二抗稀释倍数为1∶5000,最适检测范围为1-100μg/L,最小检测为0.1μg/L,批内和批间变异系数分别为3.62%和5.19%。得到回归方程(y=1.2730-0.6745x,r^2=0.9779)和标准曲线,从而建立了快速定量测定CAP含量的间接竞争酶联免疫吸附试验(ciELISA)。  相似文献   

5.
抗肠炎沙门氏菌和鼠伤寒沙门氏菌鸡卵黄抗体(IgY)的研究   总被引:1,自引:0,他引:1  
通过ELISA试验发现鸡抗肠炎沙门氏菌或抗鼠伤寒沙门氏菌卵黄抗体(IgY)具有很高的特异活性。用冻干的卵黄水溶性组分制成的肠炎沙门氏菌和鼠伤寒沙门氏菌特异性IgY粉末分别含有15.5%和10%的特异性IgY。抗肠炎沙门氏菌IgY与鼠伤寒沙门氏菌有53.3%的交叉反应,抗鼠伤寒沙门氏菌IgY与肠炎沙门氏菌有42.4%的交叉反应。曾有报道证明沙门氏菌特异性IgY可以抑制沙门氏菌在液体培养基中的生长。肠炎沙门氏菌在含有鼠伤寒沙门氏菌特异性IgY的培养基中培养4—6小时的生长速率比对照组少4倍。通过细菌计数发现鼠伤寒沙门氏菌在含有特异性IgY培养基中培养6个小时后的数量比对照组少l.6cfu/ml。用免疫荧光技术和免疫电镜技术对IgY的特异结合活性作进一步的研究,发现沙门氏菌特异性IgY能够结合沙门氏菌表面的抗原上,并导致细菌表面结构改变。  相似文献   

6.
O型口蹄疫病毒固相竞争ELISA抗体检测方法的建立   总被引:2,自引:0,他引:2  
本研究建立了O型口蹄疫病毒固相竞争ELISA抗体检测方法,确立了O型口蹄疫病毒固相竞争ELISA抗体检测方法检测O型口蹄疫血清阴阳性判定标准:抗体效价大于或等于1:32时判为O型口蹄疫抗体阳性;1:16~1:32时判为O型口蹄疫抗体可疑。检测131份阴性血清、口蹄疫亚洲1型阳性血清和A型阳性血清,固相竞争ELISA方法和液相阻断ELISA方法的特异性分别是95.5%,84.7%。固相竞争ELISA方法检测口蹄疫亚洲1型、口蹄疫A型阳性血清时,无交叉反应,O型口蹄疫病毒感染牛、免疫牛和感染猪血清的阳性检出率均为100%,免疫猪检出率为86.7%。  相似文献   

7.
用热酚-水法提取沙门氏菌O抗原,包被酶标板,建立了一种检测鸡沙门氏菌血清抗体的ELISA方法。包被抗原浓度为500ng/孔~32μg/孔,均可取得满意结果。提纯抗原与鸡的其他几种病原的抗血清不发生交叉反应。与直接凝集法(DA)比较,检测352份血清样品,ELISA阳性170份,占48%,DA阳性56份,占16%。  相似文献   

8.
为研发禽沙门氏菌抗体检测方法,采用大肠杆菌系统表达沙门氏菌属保守的菌毛fimY基因,纯化的目的蛋白经Western blot分析,能与免疫鸡血清和感染鸡血清产生特异性反应条带,证明该重组蛋白既有反应原性也有免疫原性。以此纯化蛋白为抗原建立了间接ELISA(fimY-ELISA)。通过检验已知沙门氏菌鸡源血清、相关非沙门氏菌鸡源血清对fimY-ELISA进行评价,发现其在敏感性方面高于快速平板凝集试验(RPA)和自制的脂多糖抗原ELISA(LPS-ELISA)、肠炎沙门氏菌抗原ELISA(SE-ELISA),但在特异性方面不如LPS抗原ELISA;在与RPA的符合率方面与LPS-ELISA、SE-ELISA基本一致。  相似文献   

9.
肠炎沙门氏菌 (SE)是引起人类食物中毒的重要病原之一。为了准确有效地监测鸡群肠炎沙门氏菌感染情况,日本学者建立了一种 ELISA试验,用于检测 SE特异性血清抗体。该方法包被抗原为无鞭毛的 SE抗原。试验检测血清来自某一 SE分离阳性鸡群。检测结果表明该方法的阳性率为 70%,而快速平板凝集试验 (RPA)的阳性率为 60%,以有鞭毛 SE作抗原的商品 ELISA试剂盒的阳性率为 30%。在第一次监测后约 100天,再次用三种方法检测鸡群的血样,由于 SE感染鸡变成隐性带菌鸡,因此,未分离到 SE。商品 ELISA试剂盒的阳性率为 0, RPA…  相似文献   

10.
[目的]本试验通过研究胶体金法与ELISA法在检测猪尿中盐酸克伦特罗的应用,得出两种方法在目前盐酸克伦特罗快速检测中结合使用的可能,从而在保证检测灵敏度和特异性的前提下,大大提高检测速度,并以此为基础建立一个不同于以前的监管模式。[方法]采用胶体金法和酶联免疫法(ELISA)进行比较。[结果]快速检测试纸条的灵敏度为5ng/nd,共检测442份猪尿样品,ELISA阳性7份,阳性率1.6%,胶体金阳性9例,阳性率2%;假阳性率为O.5%,符合率为99.5%;并可应用在宰后猪尿、猪肝检测中。【结论】胶体金法检测盐酸克伦特罗,具有操作简便、观察直观、快速、省时,其特异性、敏感性均达到ELISA水平;可作为饲养场、生猪中转站、屠宰场、菜市场筛查盐酸克伦特罗的重要手段,具有重要意义。  相似文献   

11.
12.
本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。  相似文献   

13.
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。  相似文献   

14.
REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.  相似文献   

15.
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.  相似文献   

16.
用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10~0.82μg/g。表明:蜂蜜应视为天然富硒营养品。  相似文献   

17.
Fractures of the anconeal process of 5 pigs ranging in age from 4 to 8 months were studied radiographically and histologically. Clinically, animals with a fracture of the anconeal process had a "tight," restricted gait. In pigs at 4.5 months of age, a radiolucent line through the base of the anconeal process was composed of fibrocartilage, fibrous connective tissue, and hyaline cartilage. Subperiosteal proliferation of woven bone was located along the cranial surface of the olecranon, adjacent to the base of the anconeal process. In older animals, the radiolucent line through the anconeal process contained variable amounts of fibrous connective tissue and fibrocartilage. The proliferation of subperiosteal bone at the base of the anconeal process formed a "buttress callus" which retained a radiolucent area between the callus and the proximal surface of the anconeal process. The latter region of radiolucency was continuous with the transversely oriented line that traversed the base of the anconeal process.  相似文献   

18.
为贯彻落实《兽药生产质量管理规范》(简称《兽药GMP》),进一步推动兽药GMP实施进程,我部制定了《兽药生产质量管理规范检查验收办法》,现予公告。本公告自2003年6月1日起施行。附件:兽药生产质量管理规范检查验收办法二○○三年四月十日第一章 总则 第一条 为推动《兽药生产质量管理规范》(以下简称兽药GMP)的实施,规范兽药GMP检查验收工作,制定本办法。 第二条 农业部负责全国兽药GMP管理和检查验收工作;负责制修订兽药GMP检查验收管理规定;负责兽药GMP检查员队伍建设和监督管理工作,负责国际兽药贸易中GMP互认工作。 …  相似文献   

19.
以国际标准强毒R株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离MG,并收集感染鸡所产蛋分离MG。结果表明,人工感染48小时后上、下呼吸道及肺已被全面感染,96小时气囊已被感染,120小时输卵管已能分离到MG,卵巢始终分离不到MG。人工感染鸡自144小时便能在其所产蛋中分离出MG。药物治疗能在72小时内消除感染,油乳剂苗则需24天后逐渐降低蛋内MG分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内MG,但以研制的种蛋浸泡剂药浴效果为最好。  相似文献   

20.
乳酸杆菌益生作用机制的研究进展   总被引:2,自引:0,他引:2  
乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和 Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。  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