合肥野生动物园5种雉鸡接种新城疫疫苗后抗体水平测定

选择成年白冠长尾雉5只,白腹锦鸡5只,白鹇3只,红腹锦鸡3只和孔雀11只,同时用L系弱毒冻干苗和新城疫油乳剂灭活疫苗进行免疫接种,通过血凝抑制试验(HI)测定其抗体滴度.结果表明,在免疫接种后26 d,白冠长尾雉抗体效价为9.8log2,白腹锦鸡为8.0log2,白鹇为8.0log2,红腹锦鸡为10.7log2;孔雀在免疫后接种0、8、15、20、25 d,抗体水平分别为3.8log2、5.0log2、6.2log2、7.5log2及8.2log2.     

BSA对牛结核抗体ELISA检测的影响

分别研究了封闭液、血清稀释液及酶标二抗稀释液中BSA成分对牛结核抗体ELISA检测的影响.结果显示,酶标二抗稀释液中BSA的存在使ELISA检测值普遍偏低,且BSA的含量越高,检测值越低;为了提高ELISA检测值,使其达到理想检测效果,必须提高酶标二抗的使用浓度.BSA中可能存在的牛血清抗体杂质对酶标二抗起到了"中和"作用,降低了酶标二抗与目标抗体结合的实际浓度,使检测结果普遍偏低.     

H5N1亚型禽流感灭活疫苗免疫鸵鸟后抗体消长规律及免疫程序的研究

为了探讨鸵鸟对H5亚型禽流感灭活疫苗的免疫原性,做好鸵鸟禽流感的防控工作,采用哈尔滨兽医研究所研制的重组H5N1亚型禽流感疫苗,不同剂量免疫鸵鸟,用HI方法检测母源抗体和免疫抗体,根据母源抗体的衰减和免疫抗体的消长规律确定首免和再免日龄.结果表明,雏鸵鸟母源抗体能维持约3周～8周;8周龄时分组首免,C1组产生的免疫反应优于C2组,抗体峰值能达到7.50 log2,维持时间为9周左右;17周龄时C1组以3.0 mL/羽进行二免,2周后抗体达到7.40log2,3周～7周抗体维持在高峰值,最高达8.80log2,以后逐渐下降,有效抗体水平能维持至接种后25周左右.二免后25周进行三免,以5 mL/羽三免后2周抗体可达到9.80log2,2周～4周抗体维持在最高峰,以后缓慢下降,期间抗体时有起伏,但有效抗体水平约可维持1年时间.三免后45周内抗体水平合格率均在70%以上.根据抗体消长规律,初步推荐了鸵鸟的免疫程序.     

Thrombolytic properties of a conjugate of liposomal urokinase with a monoclonal antibody specific for cross-linked fibrin in the model of rabbit artery thrombosis

AIM: To observe the targeting thrombolytic effect of a monoclonal antibody specific for cross-linked fibrin connected with liposomal urokinase（UK） in the model of rabbit artery thrombosis.METHODS: Preparation of thrombolytic solutions: with the method of controllable dialysis eradicator in liposomat,empty liposomes,liposomally entrapped urokinase and liposomally entrapped urokinase linked with D-dimer antibody were made.Experiment in vivo: a rabbit thrombosis model of abdominal aorta was induced by ferric chloride.When the blood pressure fall to the lowest,5 different solutions were separately imported (PBS,maximal-level UK,Ab/Lip/UK,Ab/UK-Lip,Ab-UK-Lip) and observed for 40 min continuously.RESULTS: The varieties of 5 groups blood pressures were analyzed with q-test.Significant differences were observed among Ab-UK-Lip group,maximal-level UK group and others (P<0.01).There were marked differences between Ab-UK-Lip group and others in wet weight of thrombuses (P<0.05).However,the quantity of UK in Ab-UK-Lip group was 1/3 of that in maximal-level of UK group.CONCLUSION: The thrombolytic effect on Ab-UK-Lip group is similar to that on maximal-level of UK group,but the dosage of UK is less,and the side-effect is light.This shows,as a homing-device of liposomal urokinase,the D-dimer antibody has targeting effect and the agent of Ab-UK-Lip will be attached importance by-and-by.     

Effect of different PRA serum levels from patients with β-thalassemia on the proliferation and differentiation potential of the hematopoietic stem cells /progenitor cells of cord blood

AIM: To probe the effect of different panel reactive antibody(PRA) serum levels from patients with β-thalassemia on the proliferation and differentiation potential of the hematopoietic stem /progenitor cell of cord blood.METHODS: 1×105 mononuclear cells (MNCs) isolated from umbilical cord blood were incubated with different PRA serum levels (0 μL,50 μL,100 μL) respectively and complement,inoculated into the methylcellulose cultural system.The proliferation and differentiation potential of the hematopoietic stem /progenitor cell of cord blood by the colony formation assay were detected on day 7 and day 14,respectively.RESULTS: After culture of 7 days,the total colonies and CFU-GM were 88.20±9.41,79.00±11.39 in group A and 88.60±9.12,79.20±10.44 in group B,which were significantly higher than those of 20.60±7.39,15.20±4.66 in group C and those of 4.00±2.05,1.40±0.51 in group D (P<0.01).Meanwhile compared with group A and group E,no significant difference was found (P>0.05).After culture for 14 days,the total colonies and CFU-GM were 216.00±31.10,117.40±24.80 in group A and 213.20±31.06,116.00±19.75 in group B,which were significantly higher than those of 97.80±14.43,32.80±8.10 in group C and those of 31.40±13.41,8.40±4.30 in group D (P<0.01).The CFU-GEMMs were 45.60±8.51 in group A and 42.60±7.03 in group B,which were significantly higher than those of 20.80±6.96 in group C and those of 7.80±6.06 in group D (P<0.05).The BFU-MK was 12.80±4.42 in group A and 11.00±2.74 in group B respectively,which were significantly higher than that of 1.00±0.55 in group D (P<0.05).The CFU-E in group B 17.20±4.03 was significantly higher than that of 5.60±2.87 in group D (P<0.05).Meanwhile compared with group A and group E,no significant difference was found (P>0.05).By the Kendall test,there were negative correlations between the level of PRA serum and the total colonies,CFU-GM on day 7,the total colonies,CFU-GM,CFU-GEMM,BFU-E,BFU-MK on day 14 (tau-b=-0.793,-0.849,-0.808,-0.804,-0.645,-0.674,-0.624,P<0.01).There was a negative correlation between the level of PRA serum and CFU-MK on day 14 (tau-b=-0.466，P<0.05).CONCLUSION: PRA sera inhibit the colony in the colony cultures of the hematopoietic stem cells/progenitor cells in cord blood.The inhibition depends on the level of PRA sera.The higher the level of PRA sera,the stronger the inhibition is observed in our study.     

新城疫病毒糖蛋白致病作用的研究

本实验以分子生物学和现代免疫学方法,对新城疫病毒（ＮＤＶ）中国毒株Ｆ４８Ｅ９和ＬａＳｏｔａ等毒株的糖蛋白的致病作用进行了探讨。结果表明,ＮＤＶ Ｆ和ＨＮ蛋白为糖蛋白,强弱毒株之间Ｆ蛋白裂解位点的氨基酸序列有明显的不同,并决定了毒株的毒力,但ＮＤＶ的致病作用尚需ＨＮ、Ｆ蛋白的协同作用,单一的Ｆ或ＨＮ蛋白不能构成ＮＤＶ的致病作用。     

Development and validation of an improved enzyme-linked immunosorbent assay for the detection of thyroglobulin autoantibodies in canine serum samples

An enzyme-linked immunosorbent assay to detect thyroglobulin autoantibodies (TGAB) in canine serum was developed and validated. The test result for each sample was derived from the optical density readings (OD) and expressed as an Ab-score(%) calculated from three in-house calibrators. The assay specifically detected TGAB as judged from lack of response in the assay after samples had been incubated with specific antigen. Intra- and interassay coefficients of variation ranged from 2.0–4.9% and 4.6–9.9%, respectively. The detection limit, an Ab-score of 5.6%, was close to the median Ab-score of 10% observed in healthy dogs (n = 132). The median Ab-score of dogs with primary hypothyroidism and lymphocytic thyroiditis (n = 11), skin diseases (n = 35), and non-thyroidal diseases (n = 63) was 340%, 12%, and 8%, respectively. The prevalence of TGAB in hypothyroid dogs with lymphocytic thyroiditis (sensitivity) was 91% (95% confidence limits: 59%–99%). In dogs with dermatological diseases without lymphocytic thyroiditis the prevalence of TGAB was 3% corresponding to a specificity of 97% (95% confidence limit: 85%–100%). In dogs with non-thyroidal diseases and healthy dogs the prevalence of TGAB was 5% and 6%, respectively. The diagnostic accuracy of serum TGAB was evaluated by subjecting the data from 11 dogs with lymphocytic thyroiditis and 35 control dogs without lymphocytic thyroiditis to receiver-operating characteristic curve analysis. The area under the receiver-operating characteristic curve (W = 0.966; 95% confidence limit 87%–100%) was significantly higher than that of a worthless test (0.5) (P < 0.0001), thereby indicating that serum TGAB measurements distinguished between dogs with and without lymphocytic thyroiditis.     

用大肠杆菌K99和F41菌毛抗原检测牛血清抗体的琼脂扩散试验的研究

用含２ｍｏｌ／Ｌ尿素的磷酸盐缓冲液以热处理方法提取的Ｋ９９和Ｆ４１菌毛作为抗原,对琼脂扩散试验方法进行研究。结果表明本试验方法简便,特异性强,敏感性高,可用于血清抗体检测;所制备的抗原稳定、重复性好。     

抗猪流行性腹泻病毒单克隆抗体的研制及其特性鉴定

以新生乳猪增殖猪流行性腹泻病毒（ＰＥＤＶ）,通过蔗糖密度梯度离心将其纯化,获得较为完整的病毒颗粒。病毒ＥＬＩＳＡ效价为１∶３×１０５,蛋白含量为３．９６ｇ／Ｌ。将提纯的ＰＥＤＶ免疫ＢＡＬＢ／ｃ小鼠,取免疫鼠脾细胞与ＳＰ２／０骨髓瘤细胞进行融合,融合率为９５．６％。用间接ＥＬＩＳＡ进行筛选,共检出３８个阳性孔,阳性率为２８．８％。对其中１５个阳性值较高的孔进行３次再克隆,阳性率达１００％,最后获得１５株稳定分泌特异性抗ＰＥＤＶ单抗（ＭｃＡｂ）的杂交瘤细胞。ＥＬＩＳＡ阻断试验与交叉试验证明,其分泌的抗体具有高度特异性。腹水和细胞培养上清的ＥＬＩＳＡ效价分别为１∶１０３～１∶１０６和１∶１２８～１∶５１２。经鉴定,１５株ＭｃＡｂ的分子类型均为ＩｇＧ,均无免疫沉淀特性。杂交瘤细胞的染色体数目为９０～１１０,平均９５。尤为重要的是,这些杂交瘤细胞所分泌的抗体,有的对ＰＥＤＶ具有中和作用,能够阻断病毒对猪只的侵害作用。     

马杜霉素和泰妙菌素联合应用对雏鸡新城疫抗体水平的影响

试验将不同剂量的马杜霉素和泰妙菌素分别添加到雏鸡的饲料和饮水中同时使用,通过检测不同组别雏鸡血清中ND血凝抑制效价,探讨马杜霉素和泰妙菌素不同剂量联合应用时对雏鸡免疫功能的影响。试验结果表明:正常推荐剂量的马杜霉素5mg/kg与泰妙菌素10mg/kg联合使用时,一般不会对机体免疫造成明显伤害;应用马杜霉素10mg/kg拌料能明显造成机体免疫系统伤害,当其与正常或加倍剂量的泰妙菌素联合应用更会加重对机体免疫功能的不良影响,致使抗体水平下降。