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《中国兽医学报》2017,(11)
应用PCR扩增出PEDV毒株的纤突糖蛋白S基因序列,并将其克隆到原核表达载体pGEX-4T-1中,构建出pGEX-4T-1-S表达载体。应用IPTG诱导表达重组S蛋白,并将重组蛋白以弗氏完全/不完全佐剂乳化后免疫小鼠,制备抗PEDV S蛋白的单克隆抗体,并以此建立了检测PEDV抗原的双抗体夹心ELISA方法,确定、优化了所建立的ELISA反应条件。以建立的双抗体夹心ELISA和RT-PCR方法对PEDV样品进行了比较检测,两者的符合率为100%。应用ELISA检测了吉林省疑似PED流行猪场的粪便样品,发现PEDV阳性检出率为68%。该研究为PED诊断及流行病学调查提供了一种特异、敏感、快速和易于在基层应用的技术方法及本病的防治提供了流行病学理论依据。 相似文献
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《中国兽医学报》2019,(6)
应用大肠杆菌表达及纯化的戊型肝炎病毒(hepatitis E virus,HEV)P1重组蛋白制备的抗体,建立了检测HEV抗原的双抗体夹心ELISA方法,并对吉林省和辽宁省部分地区猪群HEV的感染进行了病原流行病学调查。方阵滴定法确定鼠源捕获抗体IgG最佳包被量为0.2μg,兔源酶标抗体最佳稀释倍数为1∶500。对大量阴性粪便进行了检测与统计学分析,确定了双抗体夹心ELISA检测HEV的判定标准,即当D_(490)≥0.235判定为阳性。特异性、敏感性和重复性的试验结果表明,本试验建立的方法具有特异、敏感、快速和可重复性好等特点。该方法与RT-PCR方法相比,操作简单、便于基层推广使用。对吉林省、辽宁省不同地区猪群粪便进行检测,发现不同地区不同饲养期的猪群均存在着HEV感染,感染率为9%～33%。本试验建立的双抗体夹心ELISA方法检测HEV抗原,为HEV流行病学调查、临床诊断提供了有效的手段。 相似文献
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比较两种ELISA试剂盒检测结果的一致性,为流行病学调查和临床诊断筛选适合的商品化试剂盒。采用来自两个厂家的野毒抗体(gE)和免疫抗体(gB)ELISA检测试剂盒分别检测362份和392份猪血清,应用Kappa检验比较试验数据的一致性。结果显示:两种猪伪狂犬病抗体(g E)检测试剂盒联合检测出95份抗体阳性和256份阴性,符合率97.24%,结果一致性为极强(Kappa值0.932);两种猪伪狂犬病抗体(gB)检测试剂盒联合检测出351份抗体阳性和9份阴性,符合率91.84%,一致性为弱(Kappa值0.347)。以上结果说明,两种猪伪狂犬病抗体(gE)检测试剂盒都适用于PRV gE抗体检测,而两种猪伪狂犬病抗体(gB)检测试剂金差异较大,需慎重选择。 相似文献
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为了检查出北京市郊区某规模种猪场隐性伪狂犬病感染猪并加以淘汰,达到净化猪场的目的,应用伪狂犬病病毒(PRV)gE蛋白抗体ELISA检测试剂盒对不同阶段猪群血清进行抽检。结果表明:该种猪场不同阶段猪均存在PRV gE抗体阳性猪,采取普检种猪并淘汰PRV gE抗体阳性猪、调整免疫程序、抓好综合性防控措施、采用PRV gB蛋白抗体ELISA检测试剂盒检测免疫抗体等方法,使该种猪场猪的伪狂犬病得到了净化。说明用PRV gE蛋白抗体ELISA检测试剂盒对血清样本进行检测,发现并淘汰PRV gE抗体阳性猪、调整免疫程序可以净化猪伪狂犬病。 相似文献
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猪伪狂犬病病毒gB蛋白抗体竞争化学发光酶联免疫检测方法的建立 总被引：1，自引：1，他引：0
为了建立一种快速定量检测猪伪狂犬病病毒（PRV）gB蛋白抗体的竞争化学发光酶联免疫检测方法,以大肠杆菌表达并纯化的猪伪狂犬病病毒gB重组蛋白作为包被抗原,以辣根过氧化物酶标记的抗gB蛋白单克隆抗体作为酶标抗体,以国家参考品稀释制备的校准品绘制标准曲线实现定量检测,成功建立的PRV-gB-CLEIA在45 min内即可完成检测,可检测到最大稀释倍数为1：2 048稀释的国家参考品;与猪瘟等其他5种病毒抗原的标准阳性血清无交叉反应;批内变异系数为1.13%~9.47%,批间变异系数为2.43%~14.07%,重复性和稳定性较好。通过对采集的180份临床血清检测结果进行比较,该方法与中和试验阳性符合率为94.00%,阴性符合率为96.92%,总符合率为96.11%,明显优于商品化ELISA试剂盒。本研究建立的PRV-gB-CLEIA检测方法可以用于PRV gB抗体的快速定量检测。 相似文献
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《中国兽医学报》2017,(8):1463-1467
为了建立一种猪伪狂犬病gB抗体检测试纸,为养猪业基层工作人员提供一种快速简便的狂犬病免疫抗体评价方法。胶体金标记gB蛋白作为探针,SPA和猪抗伪狂犬病病毒多抗IgG分别作为检测线和质控线,建立了猪伪狂犬病gB抗体检测试纸。猪伪狂犬病质控血清和疫苗免疫猪血清用来评价其特异性,敏感性以及与IDEXX PRV gB ELISA试剂盒的符合率试验。试验结果显示,该试纸条具有较高的特异性和敏感性,与IDEXX PRV gB ELISA试剂盒的符合率为91.04%;该免疫层析试纸条具有快速(5min)、特异、敏感等优点,不需要专门的仪器设备和专业技术人员,适合于田间推广应用。 相似文献
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《畜牧与兽医》2014,(10):11-14
2011年以来,我国多个省份伪狂犬病免疫猪群相继暴发伪狂犬病疫情。为了探明伪狂犬病毒(PRV)流行毒株是否存在抗原变异,本研究从3个规模猪场3种PRV商品疫苗免疫的健康猪群中采集30份血清,经检测PRV gB ELISA抗体均为阳性,gE ELISA抗体均为阴性。采用PRV新流行毒株ZJ-01株和传统的LA毒株分别进行血清中和抗体测定,结果显示,3个毒株活疫苗免疫血清与ZJ-01株中和抗体效价明显低于其与LA株中和抗体效价。同时,ZJ-01株抗血清与ZJ-01株和LA株的中和抗体效价相似,其变异系数R值为0.2790.413,证明PRV分离毒株ZJ-01抗原性发生明显变异。本研究为伪狂犬病的防控提供了重要依据。 相似文献
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《畜牧兽医学报》2020,(3)
为了建立一种快速定量检测猪伪狂犬病病毒(PRV)gB蛋白抗体的竞争化学发光酶联免疫检测方法,以大肠杆菌表达并纯化的猪伪狂犬病病毒gB重组蛋白作为包被抗原,以辣根过氧化物酶标记的抗gB蛋白单克隆抗体作为酶标抗体,以国家参考品稀释制备的校准品绘制标准曲线实现定量检测,成功建立的PRV-gB-CLEIA在45 min内即可完成检测,可检测到最大稀释倍数为1∶2 048稀释的国家参考品;与猪瘟等其他5种病毒抗原的标准阳性血清无交叉反应;批内变异系数为1.13%～9.47%,批间变异系数为2.43%～14.07%,重复性和稳定性较好。通过对采集的180份临床血清检测结果进行比较,该方法与中和试验阳性符合率为94.00%,阴性符合率为96.92%,总符合率为96.11%,明显优于商品化ELISA试剂盒。本研究建立的PRV-gB-CLEIA检测方法可以用于PRV gB抗体的快速定量检测。 相似文献
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目的为某规模化猪场猪伪狂犬病病毒野毒株感染防控工作提供科学的免疫防控方案。方法对某发病猪群（采用Bartha-K61经典毒株疫苗进行了猪伪狂犬病免疫）进行流行病学调查,并采用ELISA方法检测其猪伪狂犬病gB抗体和gE抗体水平。结果根据调查、解剖和血清抗体检测结果,初步确诊该发病猪群为猪伪狂犬病野毒感染。通过采取紧急免疫伪狂犬病HB2000毒株疫苗、调整免疫程序和配合药物治疗等措施,育肥猪死亡率从2.56%降低至0.60%;除了4周龄及12周龄猪群外,其余猪群伪狂犬病野毒抗体水平全部下降;种公猪、8周龄、10周龄、14周龄猪群伪狂犬病野毒抗体阳性率均为0。结论Bartha-K61经典毒株疫苗并不能提供完全的保护力,通过紧急免疫与流行毒株同源性较高的HB2000毒株疫苗,加强生物安全管理工作,能够有效控制猪伪狂犬病野毒感染。 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