共查询到20条相似文献，搜索用时 78 毫秒
1.
犬瘟热荧光抗体技术的应用研究 总被引：16，自引：1，他引：15
用硫酸铵盐析法和蔗糖梯度离心法，从犬瘟热病毒人工感染犬的肝，脾中浓度提纯犬瘟热病毒，以其免疫家兔，制备兔抗犬瘟热病毒免疫血清，再经硫酸铵沉淀与ＱＡＥ－葡聚糖凝胶Ａ５０层析提取ＩｇＧ，于４℃低速搅拌标记异硫氰酸荧光素，制成兔抗犬瘟热病毒抗原荧光抗体。用上述荧光抗体检测２１只犬瘟热病毒人工感染犬和３５只自然感染犬的１１６份白细胞，抗原阳性１０９份，而健康犬，传染性肝炎犬，犬细小病毒肠炎犬的３７份血液白 相似文献
2.
犬瘟热病毒F1基因的原核表达与初步应用 总被引：1，自引：0，他引：1
将犬瘟热病毒(CDV)F1基因定向克隆入pET-28a( )载体．构建了原核表达质粒pET-28F1。pET-28F1转化宿主菌BL21(DE3)plysS．在IPTG的诱导下．可高效表达目的基因，表达量达菌体蛋白总量的23．52％。Western blot分析表明，所表达蛋白能与抗CDV阳性血清发生特异性的抗原-抗体反应。以E．coli表达的CDV F1蛋白为抗原．HRP标记的兔抗犬IgG为二抗．建立了检测抗CDV抗体的间接ELISA方法。试验表明．应用CDV F1蛋白作为抗原检测抗CDV抗体具有特异性高、抗原易纯化、成本低等特点。 相似文献
3.
4.
根据犬瘟热病毒（CDV）弱毒株Onderstepoort的核衣壳蛋白（NP）基因序列设计了套式引物，建立了RT-nested PCR检测方法。特异性试验表明，该法可以特异扩增出CDV NP基因片段，但从RNA病毒狂犬病病毒（RV）、犬副流感病毒（CPIV）、犬冠状病毒（CCV），DNA病毒犬细小病毒（CPV）、犬腺病毒（CAV）及正常Vero细胞中均不能扩增出条带。敏感性试验表明，RT-PCR可以扩增10^-6稀释度的病毒RNA，而建立的RT-nested PCR可以扩增到10。稀释度的病毒RNA，后者的敏感性明显高于前者。用RT-nested PCR法检测了2006年我国东北地区临床表现犬瘟热症状的病例19例，检出率达90％，明显高于RT-PCR的检出率（45％）。部分病例扩增片段的测序结果表明，CDVNP基因出现个别碱基变异。研究结果表明，建立的犬瘟热病毒RT-nested PCR方法敏感、特异，适用于动物犬瘟热的快速诊断。 相似文献
5.
荧光抗体技术在犬瘟热诊断中的应用 总被引：1，自引：0，他引：1
犬瘟热是犬的重要传染病之一。本病临床表现不一,危害严重,因此及时确诊对控制其发生和传播十分重要。本文应用犬瘟热病毒(CDV)荧光抗体(FA)技术,对某犬群流行的犬瘟热进行了诊断,现报告如下。材料和方法一、CDV荧光抗体:Bioveta Nitra公司制,工作浓度1:26。二、免抗CDV阴、阳性血清:本室制 相似文献
6.
采用常规方法建立杂交瘤细胞株并制备抗犬瘟热病毒(Canine distemper virus,CDV)(Onderstepoort株)核蛋白单克隆抗体(MAb);小鼠腹腔注射阳性克隆生产腹水;以G蛋白亲和层析法进行纯化;用透析法对单抗进行FlTC(异硫氰酸荧光素)标记;Sephadex G-25去除游离荧光素;测定F/P值及荧光抗体工作浓度;以吸收、阻断、抗原交叉试验对荧光抗体进行特异性鉴定.结果表明,获得6株能稳定分泌CDV单克隆抗体的杂交瘤细胞株,选取抗CDV核蛋白的CE3单抗成功地制备了质量较高的荧光抗体. 相似文献
7.
荧光定量RT-PCR法对犬瘟热病毒PS株感染犬血清和粪便中病毒的检测 总被引：1，自引：0，他引：1
为研究犬人工感染犬瘟热病毒(CDV)后病毒在血清和粪便中的含量及变化,本实验根据CDV核衣壳蛋白(NP)基因序列设计一对特异引物,扩增NP基因.并以NP基因重组质粒作为阳性标准品,建立检测CDV的SYBR GreenⅠ荧光定量RT-PCR方法.结果表明,该方法102拷贝～109拷贝范围内具有良好的线性关系,相关系数为R2=0.998,扩增效率为E=98.3％,敏感度高,最低检测限为6.1拷贝/μL,重复性检测变异系数低于2.62％.该方法与常规RT-PCR方法比较,其敏感性提高102倍.两只人工感染CDV PS强毒株的犬,分别于接种后17d、19d发病死亡,采用荧光定量RT-PCR方法对人工感染犬的血液和拭子液中的病毒核酸栽量进行定量检测以及对收集的22份临床样本拭子液进行检测,均检测到CDV.本研究结果表明,该方法可以用于CDV核酸定量检测,为该病毒的致病机理及病毒传播途径的研究提供了一种快速和敏感的检测手段. 相似文献
8.
9.
10.
犬瘟热重组N蛋白抗原间接ELISA方法的建立及应用 总被引：2，自引：0，他引：2
本研究以纯化的犬瘟热病毒（CDV）重组N蛋白为诊断抗原,建立了犬瘟热病毒抗体检测的间接ELISA诊断方法,将其命名为rN-CDVELISA。确定最佳抗原包被量为0．167μg/孔,待检血清最佳稀释倍数为1：100,其作用时间为45min,羊抗犬酶标抗体稀释倍数为1：2000,作用时间为60min,S／P值≥0．208者判为阳性,S／P值≤0．16者判为阴性,介于两者之间者判为可疑。该抗原不与犬细小病毒、犬传染性肝炎、犬副粘病毒、犬波特氏杆菌等4种疾病的阳性血清反应,具有良好的特异性;批内、批间重复试验,变异系数均小于15％,具有良好的重复性;检测血清样品96份,与进口诊断试剂盒的符合率为97．1％。本研究为现地免疫犬群抗体检测和进行CDV流行病学调查提供了一种简便的血清学诊断方法。 相似文献
11.
12.
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
13.
14.
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
15.
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
16.
17.
18.
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.
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