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为了研究玉米赤霉烯酮的间接竞争ELISA检测方法,试验采用牛血清白蛋白与玉米赤霉烯酮的耦联物(ZEN-BSA)做包被抗原,标准玉米赤霉烯酮(ZEN)做竞争抗原,以制备的可稳定分泌抗ZEN的单克隆抗体为基础,初步建立了ZEN间接竞争ELISA检测方法。结果表明:间接竞争ELISA检测方法线性范围为0.363 2～78.985 2μg/L,最低检测限为0.231 9μg/L;曲线回归方程为y=68.711-25.666x,其中R2=0.987 1,批内平均变异系数为3.10%,批间平均变异系数为6.26%,与相似毒素的交叉反应率均小于0.01%。说明建立的检测方法可以用于ZEN的检测。 相似文献
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在噻苯咪唑(TBZ)单克隆抗体制备基础上,建立间接竞争酶联免疫吸附法(ELISA)并组装试剂盒。结果表明,试剂盒标准曲线的线性范围为1.081.0μg/L,回归方程为Y=-2.104 8x+1.038 2(R2=0.999 8),抗体对TBZ 50%的抑制浓度为4.3μg/L;对牛奶中TBZ的最低检测限为40μg/L,牛奶样品的加标回收率范围在75.8%81.0μg/L,回归方程为Y=-2.104 8x+1.038 2(R2=0.999 8),抗体对TBZ 50%的抑制浓度为4.3μg/L;对牛奶中TBZ的最低检测限为40μg/L,牛奶样品的加标回收率范围在75.8%104.8%,批内变异系数范围为6.4%104.8%,批内变异系数范围为6.4%11.9%,批间变异系数范围为8.2%11.9%,批间变异系数范围为8.2%10.0%;TBZ试剂盒(TBZ-Kit)对TBZ特异性良好,并可在210.0%;TBZ试剂盒(TBZ-Kit)对TBZ特异性良好,并可在28℃条件下保存至少360 d。该试剂盒各项指标符合相关技术要求,具有快速、灵敏、专一、稳定的特点。 相似文献
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利用单克隆抗体间接竞争ELISA方法测定了鸡肉中环丙沙星的残留,同时利用HPLC方法进行了比较分析。间接竞争ELISA法的线性范围为6.25~400μg/L,最低检测限为4.10μg/L。HPLC法的线性范围为0.06~500μg/L,最低检测限为0.06μg/L。在标准品添加量为1.5625×10-4~1.25×10-3g/kg时,ELISA方法的平均回收率为73.81%,批内变异系数为8.75%,HPLC方法平均回收率为80.41%,批内变异系数为1.27%。结果显示,ELISA方法与HPLC方法具有很好的相关性(r2=0.9925),该ELISA方法可用于鸡肉样品中环丙沙星的检测。 相似文献
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采用直接竞争ELISA方法检测牛奶中孕酮含量.采用碳化二亚胺法,应用辣根过氧化物酶(HRP)标记11-α羟基孕酮琥珀酸酯(11α-OH-P4-HS),制备孕酮酶标抗原;酶标抗原与牛奶样品中的孕酮共同竞争结合固相包被的孕酮单克隆抗体,建立直接竞争ELISA反应体系.试验结果表明,最佳抗体包被浓度为1∶2 000,最佳酶标抗原工作浓度为1∶16 000,建立的标准曲线为y=-2.4598x+0.999(R2=0.996),牛奶中孕酮检测范围0.12～40 ng/mL,最低检测量为0.12 ng/mL,批内和批间变异系数分别为1.63%和2.49%.成功建立了可用于牛乳中孕酮快速检测的直接竞争ELISA方法. 相似文献
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《中国奶牛》2016,(3)
采用金黄色葡萄球菌肠毒素A(SEA)和金黄色葡萄球菌肠毒素B(SEB)免疫Balb/c小鼠制备相应的单克隆抗体(m Ab),并以此为基础建立SEA和SEB的ELISA检测方法,用于牛乳中金黄色葡萄球菌肠毒素的检测。SEA双抗体夹心法ELISA的曲线范围是1~16μg/L,相关系数R2=0.9996,检测牛乳的检测限是0.431μg/L,加标4μg/L和10μg/L,准确度分别为89.18%和97.62%。SEB双抗体夹心法ELISA的曲线范围是0.5~8μg/L,相关系数R2=0.9998,检测牛乳的检测限是0.335μg/L,加标4μg/L和10μg/L,准确度分别为107.83%和89.88%。 相似文献
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黄曲霉毒素B1间接竞争化学发光酶免疫分析方法的建立 总被引：1，自引：0，他引：1
本试验建立了一种用于检测玉米中黄曲霉毒素B1的间接竞争化学发光酶免疫分析方法.以辣根过氧化物酶催化鲁米诺-过氧化氢-对碘苯酚作为发光体系,通过对包被抗原、检测抗体和酶标二抗浓度的优化建立其标准曲线.所建方法的线性范围为0.31～20 μg/L,相关系数R2=0.992 7,灵敏度为0.09μg/L,处理内和处理间变异系数分别小于9.79％和13.08％,样品加标回收率为70.3％～100.85％.采用所建立的方法对10份玉米样品进行检测,检测结果与进口ELISA试剂盒的相关系数为0.831.结果表明本研究所建的方法可用于实际样品的检测. 相似文献
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黄曲霉毒素B化学发光免疫检测方法的建立 《畜牧与饲料科学》2015,36(10):29-29
在利用化学法合成吖啶酯标记的黄曲霉毒素B1(AFB1)抗原(AFB1-BSA-AE)的基础上,建立了化学发光免疫分析法(CLIA)检测样品中的黄曲霉毒素B1(AFB1)含量。结果表明,该方法对应的回归线方程为y=-16.995Ln(x)+166.06,R2=0.9996,AFB150%竞争抑制浓度(IC50)为0.924 ng/m L,检测的线性范围为0.1~5.0 ng/m L。在黍米样品中的加标回收率为75%~115%,变异系数为0.36%~1.20%。 相似文献
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将达氟沙星（danofloxacin, DFLX）与牛血清白蛋白(BSA)、卵清白蛋白(OVA)偶联分别作为免疫原与包被原, 达氟沙星为竞争的半抗原，建立间接竞争ELISA检测方法。试验结果表明,理想的包被抗原浓度为1.25 μg/L, 多抗(DFLX-PcAb)工作浓度为1∶10000,酶标二抗的工作浓度为1∶4000,最适检测范围为0.1～10 ng/L,最低检测限为0.2 ng/L, 批内和批间变异系数分别为3.81%和6.25%。得到回归方程y=0.7975-0.125x（R2=0.989）和标准曲线，从而建立了DFLX的快速检测残留的间接竞争酶联免疫吸附试验（ci-ELISA）。 相似文献
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家禽饲料中玉米赤霉烯酮化学发光酶联免疫分析方法的建立 总被引：1，自引：0，他引：1
为了建立比较间接竞争化学发光酶联免疫分析法(ic-CLEIA)与直接竞争化学发光酶联免疫分析法(dc-CLEIA)检测家禽饲料中玉米赤霉烯酮(zearalenone,ZEN)的方法,采用二因子交叉试验优化确定最佳包被原浓度和抗体稀释度,建立ic-CLEIA和dc-CLEIA。结果显示:icCLEIA在0.05~5 ng/m L范围内线性关系良好,线性回归方程为y=-0.383 5x+0.417 9(R2=0.994 6),灵敏度(IC50)为0.611 ng/m L,检出限(LOD)为15 pg/m L,平均批内和批间变异系数为1.79%和1.02%,家禽饲料添加样品回收率为91.9%~112.1%,变异系数小于10.5%;dcCLEIA在0.025~5 ng/m L范围内线性关系良好,线性回归方程为y=-0.386 5x+0.334 1(R2=0.991 8),灵敏度(IC50)为0.372 ng/m L,检出限(LOD)为3 pg/m L,平均批内和批间变异系数为1.16%和1.20%,添加样品回收率为87.3%~119.3%,变异系数小于9.8%。结果表明:两种方法精确,灵敏度高,均可作为定量检测ZEN的有效手段;与ic-CLEIA相比,dc-CLEIA更适用于检测家禽饲料中的ZEN。 相似文献
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利用制备的抗三聚氰胺单克隆抗体建立了间接竞争ELISA方法,采用棋盘滴定法确定最佳三聚氰胺-OVA包被浓度为1.0μg/mL,单抗最佳稀释度为1∶1 000,最佳的封闭液为50g/L的脱脂乳,标准曲线的线性方程为y=-15.12x+107.3(R2=0.997)。批内变异系数(CV)为1.07%～5.53%,批间CV值为3.33%～11.92%,确定此方法精确度良好。利用建立的间接竞争ELISA方法,测定添加有三聚氰胺标准溶液的样品(宠物食品、小麦面筋、饲料、牛乳、鸡蛋),得到三聚氰胺在各样品中的回收率范围分别为宠物食品73.8%～84.32%,小麦面筋67.3%～88.27%,饲料70.3%～85.9%,牛乳83.47%～89.76%,鸡蛋76.2%～87.29%。确定可以利用此间接竞争ELISA方法检测样品中的三聚氰胺含量。 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
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
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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
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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
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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
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
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