共查询到17条相似文献，搜索用时 234 毫秒
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唐成 《畜牧兽医科技信息》2018,(2)
羊附红细胞体病对羊群有一定的危害性,本文将展开对羊附红细胞体解离方法的研究。为了能够得到准确的实验结果,首先要获取红细胞感染率较高的阳性抗凝血作为实验样品,使用多种方法对红细胞体进行解离,然后比较这几种方法对羊附红细胞体的解离效果。主要采用SDS-PAGE方法检测。结果显示,在针对羊附红细胞体解离方法当中,在200m L的抗凝血液当中加入1m L的双向红莲灭,保持在4℃左右的情况下,36h后会有比较好的分离效果。传统的水溶法,其解离的效果会比较差,但可明显降低羊附红细胞的感染率及感染的强度,效率也比较高。希望能够通过本文的研究,为业界提供更多的参考。 相似文献
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从感染率90%以上的阳性牛血中分离温氏附红细胞体,利用反复冻融-超声波裂解法制备温氏附红细胞体抗原蛋白悬液,通过切胶纯化和免疫印迹法筛选具有良好免疫原性的抗原组分,制备温氏附红细胞体亚单位疫苗,并同时将抗原悬液灭活后制成温氏附红细胞体灭活苗。疫苗经检测合格后在小鼠动物模型上进行免疫持续期测定和免疫效力检验。结果显示,温氏附红细胞体亚单位疫苗对小鼠的免疫持续期为12周,免疫保护率为100%,灭活苗免疫保护率为80%,表明温氏附红细胞体特异抗原组分亚单位疫苗具有较好的免疫原性,能诱导小鼠产生良好的免疫保护作用。 相似文献
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为快速准确诊断和检测羊附红细胞体病,及时采取防治措施,建立了检测羊附红细胞体抗原的双抗夹心ELISA诊断方法。选取规模化养殖场羊,镜检附红细胞体红细胞感染率> 90%,无菌采取血液,分离羊附红细胞体抗原,制备纯化兔抗羊附红细胞体抗体,应用辣根过氧化物酶标记抗体,进行双抗体夹心ELISA试验。试验结果表明,双抗体夹心ELISA方法的最佳工作条件为:抗体最佳包被量为82.91 μg/mL,酶标抗体最适工作浓度为1∶400,抗原最低检出量为7.81 μg/mL;而且与支原体、大肠杆菌、葡萄球菌以及牛、猪、兔附红细胞体均不出现交叉反应,表明该方法具有良好的特异性,可用于羊附红细胞体病的诊断和群体检测。 相似文献
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猪附红细胞体对不同宿主红细胞的体外感染试验 总被引：1，自引：0，他引：1
为证实猪附红细胞体能否感染其它宿主红细胞,本试验在猪附红细胞体体外培养的基础上,进行了猪附红细胞体体外感染家兔、昆明小白鼠、犬、羊、牛及人的健康红细胞。结果表明,将感染猪附红细胞体的阳性血液体外感染家兔、昆明小白鼠、犬、羊、牛及人的健康红细胞,均可不同程度的感染,其中以兔和昆明小白鼠红细胞的感染率最高,分别达45.0%和40.3%;人红细胞的感染率为30.0%,呈现轻度感染;而对其它宿主红细胞,呈现一过性感染。 相似文献
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自1934年Neitz等发现并命名了羊附红细胞体(E.ovis)以来,羊附红细胞体已被公认为是致病性较强的一种病原体。目前,国内外对附红细胞体病的研究已取得了一定进展[1-2],但对羊附红细胞体抗原的研究尚少,至今尚无羊附红细胞体抗原免疫性分析的报道。试验对体外分离培养的羊附红细胞体经反复冻融、超声波裂解后,应用SDS-PAGE电泳及免疫印迹技术进行了免疫性分析,以期为羊附红细胞体病的深入研究提供依据。1材料与方法1.1材料RPM I-1640培养液(RPM IMed ium 1640)、M-199培养液(Med ium 199)、兔抗绵羊IgG-HRP,均购于北京华美公司;蛋白… 相似文献
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猪附红细胞体可溶性抗原的分析 总被引：2，自引：0，他引：2
为了解附红细胞体可溶性抗原的特性,本试验对解离下的猪附红细胞体,经超声波裂解制备粗抗原,再经SephadexG-200分离提纯后,应用对流免疫电泳(CIEP)定位特异性抗原蛋白峰,再进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)及免疫印迹(Westernblot),对猪附红细胞体粗提及纯化的可溶性抗原进行了分析。结果表明,猪附红细胞体可溶性抗原的电泳图谱上有四条蛋白带,其分子量分别为77Ku、62Ku、58Ku、29Ku,其中特异性抗原分子量为58Ku和29Ku。从而为附红细胞体抗原成分的进一步分析及该病的免疫学诊断等研究奠定了基础。 相似文献
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猪附红细胞体病的免疫学研究 总被引：5，自引：0，他引：5
将急性感染期猪全血用低渗法、皂素法、冻融法裂解红细胞经离心浓缩制备成抗原，用康复猪血清，创建了附红细胞体凝集试验，并用厌氧法创建了体外培养增殖附红细胞体的方法，用上述方法增殖浓缩抗原制成附红细胞体疫苗，临床保护期可达 8个月，抗血液寄生保护期可达 6个月，并发现早期感染猪血清中有一种免疫抑制物质。 相似文献
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以自然感染温氏附红细胞体(E.wenyoni)的牛血作为试验材料,采用镜检法观察了E.wenyoni的形态学特征,应用SDS-PAGE和免疫印迹技术分析了E.wenyoni的抗原蛋白组成。结果显示,E.wenyoni呈多形性,具有折光性,游离于血浆中的E.wenyoni具有运动性,附着于红细胞上似乎停止运动,但实际上却能使红细胞发生震颤;SDS-PAGE结果显示,E.wenyoni主要由9种蛋白组成,分子量范围为24ku～150ku,应用免疫印迹试验筛选出了3种特异性蛋白质,分子量分别为147.31ku,49.03ku和35.72ku。本试验为准确诊断牛附红细胞体感染提供了可靠依据。 相似文献
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Role of the spleen and rosette-formation response in experimental Eperythrozoon ovis infection 总被引：1，自引：0，他引：1
The role of the spleen and rosette-formation responses was investigated in sheep experimentally infected with Eperythrozoon ovis. Phagocytic activity was observed in the spleen 19 days after primary infection. Phagocytosis of E. ovis-parasitised and non-parasitised erythrocytes by cordal reticular cells occurred. E. ovis organisms seemed to be detached from the erythrocytes by pseudopodia extending from macrophages and cordal reticular cells without causing damage to the plasmalemma of the erythrocyte. No phagocytic activity was observed in spleens removed 74 and 146 days after infection. Antigen-specific lymphoid cell responsiveness, assessed by rosette formation, indicated that 2.8, 15.4, 8.0 and 6.0% of lymphoid cells in the spleens of the four E. ovis-infected sheep, respectively, formed antigen-specific rosettes. Rosette formation did not occur when splenic lymphocytes from E. ovis-infected sheep were mixed with non-infected erythrocytes or when splenic lymphocytes from an uninfected sheep were used. 相似文献
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Circulating antibody was detected by an indirect fluorescent antibody test (IFAT) in the serum of sheep infected experimentally with Eperythrozoon ovis. Antibodies were first detected 15 to 32 days after infection with E ovis and titres peaked at 41 days. This antibody may be associated, at least in part, with protection against infection with E ovis since the initial increase in antibody titre coincided with a fall in the primary parasitaemia. A role for antibody is suggested further by the fact that the prepatent period of infection was prolonged by one day and the parasitaemia initially remained at low levels in infected sheep protected by passively transferred hyperimmune serum. Moreover, following primary infection, acquired immunity was manifest by a lack of parasitaemia following challenge infections while increased IFA titres were observed. No evidence of opsonic activity was observed in an in vitro erythrophagocytosis test in that neither mouse macrophages nor sheep monocytes phagocytosed E ovis infected or uninfected erythrocytes sensitised with hyperimmune serum. 相似文献
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Effect of Eperythrozoon ovis on the lysis of sheep erythrocytes in the complement fixation test 总被引：2，自引：0，他引：2
M J Norris R S Rahaley R G Whittaker 《Veterinary immunology and immunopathology》1987,16(3-4):283-288
When erythrocytes from sheep experimentally infected with Eperythrozoon ovis were used in the titration of reagents for a standardised complement fixation test, increased amounts of both haemolysin and complement were required for erythrocyte lysis compared with preinfection titrations. The haemolysin requirement increased by up to 125% at 55 days post-infection and complement requirement increased by up to 40% at 40 days post-infection. These changes appeared to correlate with the development of a macrocytic anaemia in affected sheep rather than E. ovis parasitaemia. The results emphasise the need to carefully monitor the haematological parameters of sheep used as sources of erythrocytes for the complement fixation test. 相似文献
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分别用绵羊附红细胞体自然感染病羊的全血及分离纯化的绵羊附红细胞体对小白鼠进行攻毒,以建立绵羊附红细胞体人工感染小鼠模型。通过攻毒后症状观察、血液涂片镜检和绵羊附红细胞体特异性PCR检测法,对建立的模型进行评价。结果显示,各试验组小鼠人工感染后3~6 d,血液中均可检测到绵羊附红细胞体,而对照组小鼠未出现异常症状,且血液附红细胞体检查结果为阴性。该研究成功地构建了绵羊附红细胞体小白鼠感染模型,创建的模型可用于附红细胞体的生物学特性、致病机制、药物筛选等方面的研究。 相似文献
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Single-step purification and evaluation of recombinant BP26 protein for serological diagnosis of Brucella ovis infection in rams 总被引：1，自引：0，他引：1
Zygmunt MS Baucheron S Vizcaino N Bowden RA Cloeckaert A 《Veterinary microbiology》2002,87(3):213-220
To investigate the value of the BP26 protein in the serological diagnosis of ovine brucellosis caused by Brucella ovis, recombinant BP26 protein was produced in Echerichia coli and purified for use in an indirect enzyme-linked immunosorbent assay (I-ELISA). The majority of the recombinant protein was recovered from the supernatant of sonicated recombinant E. coli cells in a soluble form. This facilitated the purification of the recombinant BP26 protein which was achieved by using ion-exchange chromatography. After one step of purification, the purity of the recombinant BP26 protein was analyzed by using SDS-PAGE, Coomassie blue staining, and Western blot with a monoclonal antibody (MAb) directed against the BP26 protein. The degree of purity appeared satisfactory so that it could be directly used in I-ELISA. Although the recombinant BP26-ELISA appeared less useful than I-ELISA using the B. ovis hot saline (HS) extract as antigen, the high number of sera from B. ovis infected rams found positive (90%) in the recombinant BP26-I-ELISA indicated that the BP26 protein may be an additional suitable antigen for serological diagnosis of B. ovis infection in rams. 相似文献
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A modified indirect immunofluorescent assay for the detection of antibody to Eperythrozoon ovis in sheep 总被引：2，自引：0，他引：2
Experimental ovine eperythrozoonosis was studied using Giemsa staining of blood films and a modified indirect immunofluorescent antibody assay (IFAA). The serums of 21 Border Leicester Merino cross lambs between 12 weeks and 7 months-of-age were analysed before and after infection with Eperythrozoon ovis (E. ovis) using the IFAA test. No rise in the IFAA titre was seen until day 7 and this coincided with the first detection of E. ovis organisms in blood smears stained with Giemsa. The percentage of E. ovis infected red blood cells peaked on day 14, but the IFAA titre did not peak until day 35. Titres to E. ovis, on average, had begun to drop by day 63. There was considerable individual variation in response to E. ovis infection as measured by the IFAA. Titres as high as 6,400 were observed in individual sheep at the peak of E. ovis parasitaemia of red cells. One sheep had a titre of 51,200 nineteen days after infection, and titres of 3,000 were maintained for several months in a few sheep. The assay proved reliable, and up to 100 samples per day could be tested. The antigenicity of the slide preparations was found to be satisfactory after storage for 6 months at -20 degrees C and 4 degrees C and for 28 months at -70 degrees C. Temperature fluctuations during storage rendered slides unsuitable for the IFAA after these times. A method of storing E. ovis infected blood in liquid nitrogen is described. 相似文献