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1.
研究大肠杆菌热敏性肠毒素突变体蛋白(heat-labile enterotoxin,LTRG)对鸡新城疫(Newcastle disease virus,NDV)疫苗经不同途径接种效果的影响.以NDV LaSota株作为共免疫原,与LTRG重组蛋白经滴鼻、口服及肌肉注射三种途径接种雏鸡,通过血清IgG抗体、HI抗体及粘膜分泌型IgA抗体水平变化,评价不同途径接种LTRG对NDV疫苗的免疫效果的影响.结果显示:①三种途径接种,LTRG均能显著增强NDV疫苗的免疫抗体水平,免疫雏鸡血清抗体和粘膜抗体水平均高于NDV疫苗单独免疫组;②LTRG对NDV疫苗免疫增强作用,以滴鼻接种最显著:二、三免后LTRG+NDV滴鼻组与NDV组血清IgG、粘膜IgA差异均显著(P＜0.01、P＜0.05);LTRG+NDV口服与NDV组血清IgG差异不显著(P＞0.05)、粘膜IgA差异板显著(P＜0.01);LTRG+NDV肌注与NDV组IgG差异不显著(P＞0.05)、粘膜IgA差异显著(P＜0.05);③三种免疫途径比较:血清IgG抗体水平差异均不明显(P＞0.05);滴鼻、口服免疫可诱导雏鸡产生较高的粘膜IgA抗体、血清HI抗体.由此可见:LTRG粘膜佐剂对NDV疫苗免疫增强作用,以粘膜(滴鼻、口服)途径接种效果最为显著,能诱导高水平的粘膜IgA抗体及血清抗体. 相似文献
2.
以纯化后的埃博拉病毒(Ebola virus,EBOV)糖蛋白(GP)作为抗原,致敏醛化的绵羊红细胞,进行最佳反应条件优化,建立间接血凝抗体检测方法。以EBOV高免马血清、纯化的IgG和精制免疫球蛋白F(ab’)_2为待检样品进行检测,并将检测结果与假病毒中和试验检测结果相比较。结果显示,该方法可特异性检测EBOV抗体,与马尔堡病毒、裂谷热病毒等的阳性血清均不发生反应;与假病毒中和试验测得的抗体效价增长规律相一致。结果表明,本试验成功建立了EBOV抗体间接血凝检测方法,该方法安全、简便、快捷,为EBOV疫苗免疫后的效果评价提供了又一新的检测方法。 相似文献
3.
《中国兽医学报》2020,(6)
为探讨猪繁殖与呼吸综合征病毒(PRRSV)GP5和M蛋白滴鼻免疫小鼠后产生的特异性抗体的中和活性,研究以原核表达的重组蛋白GP5和M为抗原分别与霍乱毒素B亚基(CTB)佐剂按10∶1的比例混合制备成CTB黏膜佐剂疫苗,滴鼻免疫小鼠,于首次免疫后0,7,14,21,28,35,42 d收集唾液和血清样品,利用间接ELISA方法分别检测免疫小鼠唾液中特异性抗体IgA,结果均于28 d时S/N值达到最高,ELISA效价均为1∶2;血清中特异性抗体IgG分别于21,28 d时S/N值达到最高,ELISA效价均为1∶800。通过本实验室建立的基于PAM细胞中和试验方法分别检测唾液中特异性抗体IgA和血清中特异性抗体IgG中和滴度均1∶2,结果表明PRRSV GP5和M蛋白滴鼻免疫小鼠后产生的特异性抗体未达到免疫保护作用的中和抗体水平。 相似文献
4.
为研究小反刍兽疫病毒N蛋白抗体与H蛋白抗体在羊体内的代谢消长规律,试验通过建立小反刍兽疫N蛋白双抗原夹心ELISA抗体检测方法、H蛋白阻断ELISA抗体检测方法与血清中和抗体试验方法,分别检测了羊免疫疫苗后体内N蛋白抗体与H蛋白抗体在每个免疫阶段的代谢消长变化。结果表明:羊免疫小反刍兽疫疫苗后,血清内的N蛋白抗体与H蛋白抗体在6~8周时血清抗体效价较高,对应的羊体内中和抗体效价为1∶512,且效价至少可以持续到10周以上,2种抗体具有一定消长代谢规律的相关性。该研究结果也为以N抗原与H抗原为基础建立相应的抗原捕获ELISA方法、竞争ELISA检测方法、间接ELISA检测方法以及双抗原夹心ELISA检测方法奠定了一定的理论依据。 相似文献
5.
本试验应用目前国内研制的含犬副流感弱毒疫苗的五联苗,采用微量血凝与血凝抑制试验,通过对96条军犬和民犬进行定期采血检测,研究了本疫苗预防犬副流感的免疫程序,以犬副流感抗体效价达到1:64为标准,该苗的免疫程序为第一次注苗后14天再注射1次,以后每隔6个月注射1次。在将血凝与血凝抑制试验用于犬的血清副流感抗体检测中,采用了绵羊红细胞,结果清晰准确,方法有所创新。 相似文献
6.
为探讨不同免疫接种途径对鸡新城疫(ND)弱毒疫苗免疫效果的影响,我们将健康1日龄海兰蛋公鸡雏鸡(48只)随机分为滴鼻免疫组、肌肉注射免疫组、后海穴常规剂量免疫组和后海穴半剂量免疫组,每组12只.分别于第14日龄和28日龄接种新城疫IV系弱毒苗(LaSota株)1羽份/只,后海穴半剂量免疫组接种0.5羽份/只.分别于免疫前和免疫后7、14、21d和28d称重,无菌操作采血,分离血清,检测血清中的血凝抑制效价(HI).结果显示:与常规途径免疫组相比,后海穴常规剂量免疫组和后海穴半剂量免疫组可显著增强鸡血清NDV特异性HI效价(P＜0.05),且显著提高试验鸡只的日增重(P＜0.05);滴鼻免疫组与肌肉注射免疫组相比,虽然增加了HI效价,但统计学比较差异不显著(P＞0.05).试验结果说明后海穴接种鸡新城疫疫苗可显著提高鸡血清NDV的HI效价,提高鸡生产性能,同时节约疫苗用量. 相似文献
7.
《中国预防兽医学报》2017,(4)
为研究马传贫病毒(EIAV)弱毒疫苗免疫马匹后中和抗体的动态变化规律,本研究通过将表达EIAV囊膜蛋白重组质粒(pcDNA-env),与表达EIAV主要结构基因gag-pol的包装质粒(pUK-gag-pol)、表达荧光素酶的转移载体(pONY8.1)共转染293T细胞,制备了EIAV假病毒粒子(EIAV-Env)。在此基础上,利用该假病毒对2匹EIAV疫苗免疫马在免疫后6个月内不同时间点的中和抗体进行检测。即将EIAV疫苗免疫马血清通过连续4倍稀释,与EIAV-Env共孵育1 h后接种ELR1-293T细胞,36 h后通过检测细胞内荧光酶活性计算不同血清样品的中和抗体滴度。结果显示,EIAV疫苗免疫马血清可以特异性地中和EIAV-Env。EIAV疫苗免疫马后,随着时间的增加其体内中和抗体滴度逐渐升高,在6个月时能够达到最高值,这提示中和抗体可能与EIAV疫苗体液免疫成熟相关。本研究为进一步深入EIAV及其疫苗诱导体液免疫保护机制的研究提供了相关的实验依据。 相似文献
8.
本试验用犬腺病毒（ＣＡＶ）Ⅱ型狐狸脑炎疫苗免疫接种２－２．５月龄离乳仔狐，分别用血凝抑制试验（ＨＩ），补体试验（ＣＦ），中和（ＮＴ）检测免疫后不同时期狐狸脑炎抗体滴度，结果表明免疫后７天三种抗体均出现滴度，２１－３０天达到高峰，直至屠宰前保持不降（中和抗体滴度略有下降）。站体结合试验操作简单，重复性好，特异性高，结果可靠等优点，适合大批狐狸脑炎抗体的检测。 相似文献
9.
不同种禽类红细胞悬液对HI结果的影响 总被引：5，自引：0，他引：5
《养禽与禽病防治》2005,(1)
红细胞凝集抑制试验能够用于监测某些畜禽传染病,如新城疫、禽流感等的抗体水平,由于动物血清中存在某些非特异性血凝物质,致使血凝抑制试验的结果出现非特异性血凝现象。本实验采用鸡、鸭和鹅的红细胞分别与同一份血清进行血凝抑制实验,结果表明,用鸡红细胞来检测鸭、鹅血清抗体效价时,出现非特异性血凝的血清份数较少;而用鸭、鹅红细胞来检测鸡血清的抗体效价时,出现非特异性血凝的血清份数较多。 相似文献
10.
为建立一种快速的新城疫病毒抗体检测方法,本研究以原核表达的重组HN蛋白作为诊断抗原,建立了检测新城疫病毒抗体的间接ELISA诊断方法.该方法检测AIV(H9亚型)、IBV、IBDV、EDSV 4种常见禽病病原的阳性血清均为阴性;检测灵敏度为1:12 800;批内重复性试验、批间重复性试验的变异系数分别小于5％和10％;与血凝抑制试验(HI)符合率为96.1％.本研究建立的NDV HN间接ELISA检测方法具有良好的特异性、敏感性和重复性,为NDV的抗体检测及流行病学调查等快速诊断提供一种技术手段. 相似文献
11.
SIMULATED NATURAL INFECTION OF CHICKENS WITH AUSTRALIAN LENTOGENIC NEWCASTLE DISEASE VIRUS AND SUBSEQUENT CHALLENGE WITH VIRULENT VIRUS 总被引：1，自引：0，他引：1
A. J. Turner M.V.Sc. Ph.D. R. P. Hanson Ph.D. J. Spalatin V.M.D. 《Australian veterinary journal》1976,52(11):524-528
A total of 291 eight-week-old chickens were exposed to chickens infected with either of two Australian lentogenic strains (V4 and AVL NDV-1) of Newcastle disease virus (NDV). At 3 weeks after exposure, all chickens exposed to V4 infected chickens had developed haemagglutination-inhibition (HI) antibody. All chickens exposed to AVL NDV-1 virus infected chickens had developed HI antibody 5 weeks later. This sudden late appearance of HI antibody, to titres higher than those observed with V4 chickens, was explained by V4 virus being introduced to the AVL NDV-1 group of chickens. When groups of these chickens were challenged with Roakin virus (mesogenic NDV) at 3 weeks and Fontana 1083 virus (viscerotropic velogenic NDV) and Texas GB virus (neutrotropic NDV) at 3, 5, 10 and 21 weeks only three chickens developed clinical illness one of which died. These chickens were one AVL NDV-1 chicken contact challenged with Fontana 1083 virus at 3 weeks, one V4 chicken oronasally challenged with Texas GB virus at 5 weeks and one V4 chicken challenged oronasally with Fontana 1083 virus at 10 weeks. Susceptible non-vaccinated chickens died soon after challenge. Challenge by oronasal infection with 10(7.0) ELD50 of virus or contact with susceptible infected chickens enabled virulent virus to be isolated from most chickens and was accompanied by a large anamnestic increase in serum HI antibody. 相似文献
12.
H Mészsáros 《DTW. Deutsche tier?rztliche Wochenschrift》1991,98(4):123-126
The NDV-6-strain was selected by Lomniczi from naturally occurring Newcastle-disease-virus-strains. This strain was administered to chicken of different ages without detrimental effects. The strain was thermoresistant and adapted by serial passages in gut epithelial cells (NDV-6/10 variant). By using an aerosol of this strain (10(7),(3) EID50/m3) more than 1,3 millions chicken were immunised either in the incubator and up to an age of 10 days. Vaccine reactions were not observed. Vaccination against Marek's disease did not interfere with NDV-vaccination efficacy when day-old chicken were vaccinated with the NDV-6/10 aerosol. Obviously, the vaccine elicits a distinct cell-associated immunity. This explains, why chicken exhibiting low hemagglutination inhibition serum-antibody titers were found resistant to challenge infection with velogenic NDV. Immunity of chicken which have been vaccinated within 1 to 8 days post hatch by the NDV-6/10 aerosol vaccine, persisted 6 to 8 weeks and can be extended by revaccination up to onset of laying. The vaccine is now commercially available and designated VITAPEST. 相似文献
13.
Ramp K Veits J Deckers D Rudolf M Grund C Mettenleiter TC Römer-Oberdörfer A 《Avian diseases》2011,55(3):413-421
To analyze the contribution of neuraminidase (NA) toward protection against avian influenza virus (AIV) infection, three different recombinant Newcastle disease viruses (NDVs) expressing hemagglutinin (HA) or NA, or both, of highly pathogenic avian influenza virus (HPAIV) were generated. The lentogenic NDV Clone 30 was used as backbone for the insertion of HA of HPAIV strain A/chicken/Vietnam/P41/05 (H5N1) and NA of HPAIV strain A/duck/Vietnam/TG24-01/05 (H5N1). The HA was inserted between the genes encoding NDV phosphoprotein (P) and matrixprotein (M), and the NA was inserted between the fusion (F) and hemagglutinin-neuraminidase protein (HN) genes, resulting in NDVH5VmPMN1FHN. Two additional recombinants were constructed carrying the HA gene between the NDV P and M genes (NDVH5VmPM) or the NA between F and HN (NDVN1FHN). All recombinants replicated well and stably expressed the HA gene, the NA gene, or both. Chickens immunized with NDVH5VmPMN1FHN or NDVH5VmPM were protected against two different HPAIV H5N1 and also against HPAIV H5N2. In contrast, immunization of chickens with NDVN1FHN induced NDV- and AIV N1-specific antibodies but did not protect the animals against a lethal dose of HPAIV H5N1. Furthermore, expression of AIV N1, in addition to AIV H5 by NDV, did not increase protection against HPAIV H5N1. 相似文献
14.
为了解中国鸭源新城疫病毒(Newcastle disease virus,NDV)的毒力特点,从2009年广东地区发病鸭群中分离和鉴定出1株新城疫病毒(简称NDV-104),对其生物学特性、致病性和融合蛋白(fusion,F)、血凝素神经氨酸酶(hemagglutinin-neuraminidase,HN)基因进行了研究。结果显示,分离株的MDT、ICPI和IVPI分别为56.4 h、1.95和1.64,结合F蛋白裂解位点(112~117位)的氨基酸序列分析,确定了分离株为新城疫强毒。致病性结果表明,分离病毒对雏鸭具有感染性和致病性。F基因遗传进化结果显示,分离株属于基因Ⅶd亚型。F、HN蛋白的氨基酸同源性结果表明,分离株与2000年以来国内外分离到的基因Ⅶ型NDV同源性较高,分别为96.6%~99.3%和97.0%~99.7%,而其与常用疫苗株B1、V4、Clone30、Mukteswar和LaSota的F、HN蛋白氨基酸序列的同源性较低,且与LaSota株和V4株的同源性最低,分别仅为88.1%和87.6%,说明分离株与经典新城疫病毒毒株存在一定的差异。 相似文献
15.
新城疫病毒和禽流感病毒复合型胶体金免疫层析试纸条的制备 总被引：4，自引：0，他引：4
为了建立一种简便、快速而且能同时检测新城疫病毒(NDV)和禽流感病毒(AIV)的方法,本试验采用柠檬酸三钠还原法制备胶体金颗粒,分别标记NDV单克隆抗体6C4和AIV单克隆抗体制备免疫检测探针。在硝酸纤维素膜上,用微定量喷头喷好2条病毒检测线(T线)和1条羊抗鼠抗体质控线(C线),制备复合型免疫层析检测试纸条。结果在10 min内,可同时检测出两种病毒。试纸条检测NDV的灵敏度比HA试验结果提高8倍;AIV重组抗原的检测灵敏度为1.7μg/mL。两种病毒互相测试,未出现交叉反应。用缓冲液对照测试结果为阴性。在密封、干燥、低温的条件下,试纸条的灵敏性与特异性没有明显变化。说明本研究建立的NDV和AIV免疫层析检测法具有特异、灵敏、稳定、操作简单等特点,符合现场快速检测的要求。 相似文献
16.
17.
The avirulent Newcastle disease virus strain designated NDV-6/10, selected by B. Lomniczi at the Veterinary Medical Research Institute, Hungarian Academy of Sciences, is completely safe for day-old chickens by aerosol vaccination. Aerosol immunization using the Hungarian-made MASTERDROP generator (particle size: maximum 7 microns) caused no vaccination reactions among 206,000 chickens with different maternal antibody levels. Other vaccines given simultaneously did not significantly affect the protection elicited against Newcastle disease (ND). Almost 100% and 90% of the aerosolized chickens survived subcutaneous challenge with 10(6) LD50 NDV at 30 and 50 days old, respectively. A single immunization is sufficient for broilers; however, parent flocks should be revaccinated at 7 so 8 weeks old. 相似文献
18.
The effects of viral vaccinations and immunization with sheep red blood cells (SRBC) on the humoral response of pullets were investigated. Pullets were vaccinated with Marek's disease virus, Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious bursal disease virus at appropriate ages used in commercial practice. At seven weeks, the pullets were intramuscularly immunized with SRBC. NDV and IBV antibodies were detected by hemagglutination-inhibition tests. Hemagglutination (HA) titers were established against SRBC. IBV antibody titers were not affected by vaccination or by immunization with SRBC. NDV antibody titers were significantly increased by vaccination and by immunization with SRBC. The SRBC agglutinin response was also positively affected by vaccination. The HA titer increase consisted of a rise in 2-mercaptoethanol (2-ME)-sensitive antibodies and a fall in 2-ME-resistant antibodies. 相似文献
19.
Sakai K Yada K Sakabe G Tani O Miyaji K Nakamura M Takehara K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(5):491-494
Serum samples from 191 ostriches (Struthio camelus) in Japan were tested for antibodies to Newcastle disease virus (NDV) and avian influenza virus (AIV). Twenty-two (12%) contained NDV-specific neutralizing antibodies by a virus-neutralization (VN) test without vaccination. Antibodies to AIV were not detected in the any sera by an agar gel precipitation test. Seven serum samples that had vaccinated with live NDV by eye drop were all positive by the VN test at 1 month post vaccination. A haemagglutination inhibition (HI) test for NDV seemed not to be suitable for ostriches because of non-specific agglutination of chicken red blood cells. No haemagglutinating viruses were isolated. This is the first report on detection of antibodies against NDV in ostriches in Japan. 相似文献
20.
新城疫病毒胶体金免疫层析试纸条的制备 总被引：1，自引：1，他引：0
为了建立一种简便、快速检测新城疫病毒（Newcastle disease virus, NDV）的方法,本试验采用柠檬酸三钠还原法制备胶体金颗粒,标记NDV单克隆抗体6C4制备免疫检测探针。在硝酸纤维素膜上,用微定量喷头喷好1条病毒检测线（T线）和1条羊抗鼠抗体质控线（C线）,制备免疫层析检测试纸条,使用该制备好的检测试纸条可在10 min内检测出新城疫病毒。试纸条检测NDV的灵敏度比HA试验结果提高8倍,与禽流感病毒未出现交叉反应,用缓冲液对照测试,结果为阴性,在密封、干燥、低温的条件下,试纸条的灵敏性与特异性没有明显变化。本研究建立的NDV免疫层析检测法具有特异、灵敏、稳定、操作简单等特点,符合现场快速检测的要求。 相似文献