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斑点免疫金渗滤法检测猪瘟抗体的研究 总被引:14,自引:0,他引:14
以猪瘟抗原包被硝酸纤维素膜,然后用胶体金标记SPA,建立检测猪瘟抗体水平的斑点免疫金渗滤法检测试纸盒。通过胶体金标记金黄色葡萄球菌A蛋白(SPA)直接显色,阳性者出现红色斑点,结果易于判断。整个试验过程仅需5min,操作简单,与猪兰耳病、猪伪狂犬病、猪细小病毒、新城疫和禽流感等阳性血清不发生交叉反应。同时将该法与目前猪瘟的常规检测方法Dot—ELISA法和间接血凝试验同时对200份猪血清进行猪瘟抗体检测比较,符合率达98.4%和98.9%。说明该法微量、特异、敏感可靠,检测时间短,效果直观,非常适用于猪瘟的早期诊断和普查以及疫苗免疫效果后抗体水平的检测。 相似文献
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猪伪狂犬病血清抗体gE-ELISA检测方法的建立 总被引:4,自引:0,他引:4
以纯化的猪伪狂犬病病毒gE蛋白为抗原建立了检测猪伪狂犬病血清抗体的间接gE-ELISA方法。最佳反应条件为,抗原包被浓度为1.7μg/mL,待检测血清1:40稀释。与伪狂犬病阳性血清反应为阳性,与猪瘟、猪细小病毒病、猪繁殖与呼吸综合征(猪蓝耳病)、猪乙型脑炎、猪布氏杆菌病5种疾病阳性血清和猪伪狂犬病gE缺失疫苗接种的猪免疫血清及SPF猪阴性血清均无交叉反应。批间、批内试验变异系数分别不超过5%和9%。用该方法与Ingezim ELISA试剂盒和HerdChek ELISA试剂盒同时对172份血清进行了平行检测,总符合率分别达93.6%和83.7%。试验结果表明:猪伪狂犬病血清抗体间接gE-ELISA检测方法具有较高的敏感性和特异性,且重复性好,可用于猪伪狂犬病野毒感染猪的血清抗体检测。 相似文献
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斑点免疫金渗滤法检测猪生殖与呼吸综合征抗体的研究 总被引:4,自引:0,他引:4
应用提纯的猪生殖与呼吸综合征病毒(PRRSV)抗原包被硝酸纤维素膜,然后用胶体金标记的金黄色葡萄球菌A蛋白(SPA)建立了检测PRRS抗体水平的斑点免疫金渗滤法(DIGFA)。该法通过胶体金标记SPA直接显色,阳性者出现红色斑点,整个试验过程仅需5min即可判断结果;与猪瘟、猪伪狂犬病、猪细小病毒病阳性血清不发生交叉反应;纯化PRRSV抗原的最低检测量为0.0553mg/mL,即55.3ng/点。对200份猪血清用该法与ELISA同时进行PRRS抗体检测,两种方法的符合率达98%。 相似文献
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为了解猪伪狂犬病在闽北地区的流行情况,整理并分析2015年度血清抗体检测数据。采用ELISA检测方法对闽北地区45个规模化猪场进行检测分析,通过检测样品血清中伪狂犬病gE抗体和gB抗体的水平进行结果判定。结果显示:送检的45个规模化猪场,伪狂犬病gE抗体阳性场达25个,猪场阳性率达55.6%;共检测1304份血清,其中伪狂犬病gE抗体阳性213份、可疑24份、阴性1067份,抗体阳性率达20.6%。从伪狂犬病gE抗体阴性猪场抽检猪伪狂犬gB抗体380份,gB抗体阳性258份,猪群伪狂犬病抗体保护率仅有67.9%。 相似文献
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间接ELISA检测猪伪狂犬病血清抗体 总被引:9,自引:0,他引:9
用猪肾传代细胞IBRS-2增殖猪伪狂犬病病毒(PRV)鄂A株,病毒培养上清液经硫酸铵沉淀、聚乙二醇(Mr20000)浓缩后作为包被抗原。用纯化的猪血清IgG免疫家兔,HRP村记撮的兔抗猪IgG,制备出高效价的酶标抗体,酶标抗体工作浓度为1:50000;经各种条件的选择,建立了检测猪伪狂犬病血清抗体的间接ELISA。所建立的间接ELISA抗原包被浓度为39.2mg/L,血清最佳稀释度为1:20,与猪细小病毒、猪、O型口蹄疫、猪衣原体标准阳性血清呈阴性反应,与标准阴性血清和临床未感染PRV的猪血清呈阴性反应;与猪伪狂犬病标准阳性血清、免疫猪血清和临床发病猪血清呈明显的阳性反应;与美国进口的PRV抗体检测ELISA诊断试剂盒检测结果比较,45份猪血清的阴、阳性检出符合率均为100%。表明建立的间接ELISA具有敏感性高、特异性强、重复性好的优点,可用于猪伪狂犬病血清抗体的定性和定量检测。 相似文献
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《中国兽医杂志》2016,(9)
为建立一种简单、快速、敏感、特异的猪伪狂犬病病毒野毒抗体血清学检测方法,本研究利用胶体金标记SPA蛋白作为金标垫,以重组g E蛋白和猪Ig G分别作为检测线和质控线,组装检测猪伪狂犬病病毒g E抗体的胶体金免疫层析试纸条。该试纸条肉眼于15 min内即可判定结果,检测PCV-2、PRRSV、PEDV、CSFV、PPV、PRV疫苗毒的阳性血清均为阴性,检测PRV野毒阳性血清的灵敏度达1∶1 280,与IDEXX g E-ELISA抗体检测试剂盒的符合率为95.31%,室温条件下可稳定保存6个月以上。本研究研制的PRV野毒抗体胶体金免疫层析试纸条操作简单、检测快速、敏感性高、特异性强,可用于PRV野毒感染的快速诊断,特别适合于现场检测。 相似文献
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《国外畜牧学(猪与禽)》2021,(5)
为了解猪伪狂犬病在规模化猪场的净化效果,采用ELISA对规模化猪场血清检测样品中猪伪狂犬病病毒gE抗体的水平进行判定。结果显示:净化前,3个规模化猪场共检测34份血清样品,其中猪伪狂犬病病毒g E抗体阳性3份,抗体阳性率达8.8%。采取净化措施后,3个猪场共检测34份血清样品,未检出猪伪狂犬病病毒gE阳性抗体,净化措施取得明显的效果。 相似文献
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应用胶体金免疫层析技术,结合胶体金定量读数仪研制出一种快速定量检测猪血清中猪瘟抗体水平的检测卡。该检测卡以胶体金标记高灵敏的猪瘟重组E2蛋白,同时在NC膜上包被同一蛋白,经正交实验确定最适条件,同时通过与对照线的颜色对比,应用胶体金定量读数仪,可对样本中猪瘟抗体水平进行定量。该检测卡检测方法简便、快速、稳定性好。与猪圆环病毒病、猪伪狂犬病、猪繁殖与呼吸综合征等常见猪疫病抗体均无交叉反应。检测71份猪血清样本结果与ELISA结果的符合率达90.1%。结果表明,实验研制的猪瘟抗体胶体金快速定量检测卡可用于基层兽医站和养殖企业的检测。 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