传染性法氏囊病的防治

目的 研究以传染性法氏囊病病毒制成多价弱毒疫苗进行IBD的免疫预防效果。方法 分别从江苏、浙江等8省(市)采集IBD病鸡法氏囊病料,分离到32个野毒株,经SDS-PAGE鉴定,其中17株病毒为IBDV。应用交叉中和试验,将分离的IBDV分为5个亚型。用IBDVI型变异株高免血清作中和试验,其中5株有较高的中和效价,初步定为IBDVI型变异株。将其在CEF上连续传代致弱,与标准I型弱毒株联合制成多价弱毒疫苗。结果 试验表明它具有良好的免疫效果,免疫组攻毒100％保护,对照组攻毒死亡率为92％。以IBD病变典型的法氏囊组织制成的IBD油乳剂灭活疫苗具有满意的免疫效果。结论 将之免疫健康鸡和IBD康复鸡,制备高免血清和卵黄抗体,血清抗体的AGP效价均达1:32,最高达1:128,治疗IBD病鸡的有效率达90％～98％;卵黄抗体的AGP效价达1:64,应用于IBD病鸡治疗,治愈率达90％～95％。制备的二联卵黄抗体用于治疗IBD和ND混合感染病鸡,治愈率达71％。     

鸡传染性支气管炎的研究——Ⅱ.利用鸡传染性支气管炎病毒在鸡胚内对B_1系新城疫病毒的干扰作用诊断鸡传染性支气管炎

本试验利用鸡传染性支气管炎病毒(IBV)在鸡胚内对B_1株新城疫病毒(NDV—B_1)的干扰现象作为鸡传染性支气管炎(IB)的一种诊断方法。对其特异性、敏感性、操作方法以及对天然病例的诊断效果等几个方面的问题进行了探讨。 用IBV强毒人工发病的病鸡组织上清液接种鸡胚后10小时,再接入一定量的NDV—B_1,则50％以上鸡胚的尿囊液的HA滴度在1:20以下,而仅接NDV—B_1的对照组中,90％以上鸡胚的尿囊液的HA滴度在1:40或1:40以上,说明IBV在鸡胚内明显地干扰了NDV—B_1,在IBV被相应的抗血清中和之后,这种干扰作用也随之消失。 新城疫病毒(NDV)、鸡传染性喉气管炎病毒(ILTV)、鸡痘病毒(FPV)、鸡败血霉形体(MG)等鸡呼吸道病的病原在鸡胚内对NDV—B_1则没有这种干扰现象。 利用这种干扰现象对6个鸡场的呼吸道病的病例进行诊断试验,结果4个鸡场为IB阳性,2个鸡场为IB阴性,这一结果与用病料在鸡胚内进行盲传继代及小鸡人工发病的结果是相符的。 试验证明,IBV在鸡胚内对NDV—B_1的干扰试验可以作为IB的一种诊断方法。     

系统性红斑狼疮47例中六种抗体阳性率比较

目的评价抗核小体抗体(Anua)、抗核抗体(ANA)、抗双链DNA(ds-DNA)抗体、抗Sm抗体、抗组蛋白抗体(AHA)、抗SSA抗体6种抗体对系统性红斑狼疮(SLE)诊断的敏感性。方法选取我院2008年10月至2010年12月在我院风湿免疫科住院SLE患者及门诊健康体检者,用酶联免疫吸附试验(ELISA)法检Anua和AHA水平,用间接免疫荧光检测ANA和抗ds-DNA抗体,用酶联免疫斑点法检测抗Sm抗体和抗SSA抗体,并用免疫双扩散法证实抗Sm抗体和抗SSA抗体的结果。结果 Anua阳性率为77%,AHA阳性率为43%,抗ds-DNA抗体阳性率为32%,抗Sm抗体阳性率为28%,ANA阳性率为94%,抗SSA抗体阳性率为17%。结论 Anua的阳性率明显高于抗ds-DNA抗体和抗Sm抗体阳性率可能是诊断SLE比抗ds-DNA抗体更特异的指标,尤其对于抗ds-DNA抗体阴性的SLE患者的诊断更具有临床应用价值;抗ds-DNA抗体和抗Sm抗体,这2种抗体对诊断SLE特异性高,但因其灵敏度低,故阳性率不高;ANA因阳性率高,而特异性低,只适宜作为SLE的筛选试验;SSA抗体阳性率在6种抗体中最低,不适宜作为筛查项目,但其真阴性率高。因此6种抗体联合检测可互相弥补其缺陷,有利于SLE诊断及治疗。     

免疫复合物用于免疫防御病毒性疾病

本文首次报告了新城疫病毒(NDV)抗原亚单位与特异性多克隆同种抗体组成的免疫复合物(IC)对鸡的保护作用。 在NDV感染的鸡胚尿囊液中加入最终浓度为0.2％的Triton X-100,37℃作用24小时后作超声处理,并离心澄清,即制成     

抗病毒口服液在鸡胚内的抗病毒作用研究

目的观察抗病毒口服液在鸡胚内的抗病毒作用.方法采用鸡胚法(体内及体外)研究了抗病毒口服液对三种病毒(甲3型流行性感冒病毒IFV株、亚甲型流行性感冒病毒鼠适应的FM1株及新城疫鸡瘟病毒NDVⅡ系病毒株)的作用.结果抗病毒口服液对三种病毒株在鸡胚体外有明显的抑制作用或杀灭作用,在鸡胚内对鸡胚尿囊液HA效价有明显抑制趋势,但统计学处理无明显差异.结论抗病毒口服液在体外对甲3型流感病毒株、亚甲型FM1株和NDVⅡ系病毒株有明显的抑制作用,抗甲3型流感病毒和亚甲型FM1株的作用比抗NDVⅡ系病毒株的作用略强,但该药在鸡胚内抗病毒作用仅有抑制趋势.     

抗白色念珠菌IgY制备及其防治小鼠感染的研究

目的：观察白色念珠菌(白念菌)免疫产蛋鸡后制备的抗白念菌蛋黄IgG抗体(IgY)的性状，探讨其体内生物学效应。方法：(1)应用白念菌为抗原免疫25周龄Leghorn鸡，改良水溶法(WD)提取蛋黄抗体IgY，双紫外光测定抗体含量，SDS—PA(江电泳检测抗体纯度，Western blot测定抗体IgY的性状。(2)建立烧伤SPF小鼠白念菌感染模型，活菌计数法检测回肠黏膜黏附白念菌量及盲肠内容物白念菌量，ELISA检测血浆TNFα和二胺氧化酶(DAO)含量。结果：(1)提取抗白念菌IgY抗体含量13．7g／L蛋黄液，抗体纯度95％，该抗体与鸡血清中IgG具有相同相对分子质量和抗原性。(2)烧伤SPF小鼠喂服IgY后，能明显抑制肠道内白念菌生长及其黏附肠上皮细胞的能力；明显降低血浆中DAO和TNFα含量。结论：WD水溶法可得到蛋黄内高产量、高纯度的特异性抗体IgY，而且具有良好的体内生物学效应。     

输血前传染性感染疾病640例检测结果分析

目的:分析输血前乙型肝炎、丙型肝炎、梅毒和艾滋病的检测结果.方法:对我院640例受血者用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELASA)进行输血前乙型肝炎病毒表面抗原(hepatitis B rirus surface antigen,HBsAg)、丙型肝炎病毒抗体(抗-HCV)、梅毒抗体(抗-TP)、人类免疫缺陷病毒抗体进行检测分析.结果:HBsAg阳性75例(11.7％(,抗-HCV阳性10例(占1.56％);抗-HIV1+2型阳性1例(0.15％);抗-TP阳性3例(占0.47％);输血前四项总阳性89例(13.9％).结论:对输血前进行感染疾病的检测,可以了解受血者输血前状况,发现潜伏期无症状感染,为诊断和治疗提供依据和帮助.     

抗眼镜王蛇毒鸡卵黄抗体的制备

目的　研制抗眼镜王蛇毒鸡卵黄抗体 ,并研究该抗体在鸡卵黄中表达过程。方法　以甲醛减毒处理的眼镜王蛇粗毒免疫莱航母鸡 ,母鸡免疫应答后 ,在卵黄中产生抗眼镜王蛇毒抗体 ,应用嗜硫色谱柱HiTrapIgYPurificationHP从鸡卵黄中提取抗眼镜王蛇毒抗体 ;以间接ELISA法及双向免疫抗散法测定抗体的效价、特异性及免疫活性 ;采用福林酚法测定蛋白含量 ;采用SDS PAGE法测定分子量及鉴定抗体纯度 ;抗体的特异性及交叉反应采用酶联免疫吸附法进行进行验证。结果　母鸡初次免疫第 9天即有抗体产生 ,初次免疫 6 0天后抗体效价超过 10 5;卵黄抗体经嗜硫色谱柱纯化后 ,经SDS PAGE检测为电泳纯 ,纯化后抗体效价较纯化前提高 4倍 ,每枚蛋中平均可获得较纯抗体 97 5mg ;所获取的抗体具有高度特异性 ,与蝰蛇科属的其它蛇毒无交叉反应 ,与眼镜蛇科蛇毒存在不同程度交叉反应 (蛇毒浓度大于5 0 0 μg·L-1时明显 )。结论　本研究首次研制并鉴定了抗眼镜王蛇毒鸡卵黄抗体 ,开拓了我国蛇伤治疗的新领域 ,并分析抗体在产蛋期卵黄液中表达的进度 ,为今后该抗体的持续、高质量诱导奠定基础 ,同时也促进其它抗蛇毒鸡卵黄抗体的研究开发及蛇毒单克隆抗体的研制。     

抗核抗体对不同抗原的免疫应答

目的　探讨体内产生抗核抗体的抗原。方法　分别以羊红细胞、伤寒沙门菌及鸡血清免疫小鼠 ,同时设立生理盐水对照组 ,每组 10只 ,采用商品化酶联免疫法检测抗核抗体试剂盒 ,将酶结合物 (酶标抗人IgG)换成酶标金黄色葡萄球菌A蛋白 (SPA) ,建立检测抗核抗体的SPA ELISA法 ,并使其性能保持不变 ,用其检测小鼠的抗核抗体。结果　新建立的检测抗核抗体的SPA ELISA法与原法检测同一批标本 ,相关系数为 0 931,P <0 0 1。小鼠血清抗核抗体检测结果 (阳性率 ) :羊红细胞组为 90 % (9 10 ) ,伤寒沙门菌为 70 % (7 10 ) ,鸡血清组为 10 0 % (10 10 ) ,生理盐水对照组未检出抗核抗体。各实验组与对照组比较差异有显著性 (P <0 0 1)。结论　体内抗核抗体可能是一类由不同抗原刺激非特异产生的抗体。     

类风湿关节炎患者血清抗巨细胞病毒抗体的临床应用

类风湿关节炎（RA）是一种常见的风湿性疾病,病因不详。目前认为其发病与遗传、免疫及环境因素有关。在环境因素中,病毒可能是RA发病的一种触发因素[1],其中巨细胞病毒在RA发病中的作用一直是个有争议的问题,认为巨细胞病毒与RA的发病有关。本研究应用酶联免疫法（ELISA）首次报道RA血清中抗巨细胞病毒IgM抗体,报告如下。     

Quaternized chitosan nanoparticles loaded with the combined attenuated live vaccine against Newcastle disease and infectious bronchitis elicit immune response in chicken after intranasal administration

Newcastle disease (ND) and infectious bronchitis (IB) are important diseases, which cause respiratory diseases in chickens, resulting in severely economic losses in the poultry industry. In this study, N-2-hydroxypropyl trimethyl ammonium chloride chitosan (N-2-HACC) and N,O-carboxymethyl chitosan (CMC) were synthesized as adjuvant and delivery carrier for vaccine antigens. N-2-HACC-CMC/NDV/IBV nanoparticles (NPs) (NDV/La Sota and IBV/H120 encapsulated in N-2-HACC-CMC NPs) and N-2-HACC-CMC/NDV-IBV NPs (the mixing of N-2-HACC-CMC/NDV NPs and N-2-HACC-CMC/IBV NPs in a ratio of 1:1) were prepared by the polyelectrolyte composite method, respectively. Both nanoparticles exhibited lower cytotoxicity and higher stability. Their bioactivities were maintained when they were stored at 37?°C for three weeks. Release assay in vitro showed that both NDV and IBV could be sustainably released from the nanoparticles after an initial burst release. In vivo immunization of chickens showed that N-2-HACC-CMC/NDV/IBV NPs or N-2-HACC-CMC/NDV-IBV NPs intranasally induced higher titers of IgG and IgA antibodies, significantly promoted proliferation of lymphocytes and induced higher levels of interleukine-2 (IL-2), IL-4 and interferon-γ (IFN-γ) than the commercially combined attenuated live vaccine did. This is the first study in the field of animal vaccines demonstrating that intranasal administration of chickens with antigens (NDV and IBV) encapsulated with chitosan derivative could induce humoral, cellular, and mucosal immune responses, which protected chickens from the infection of highly virulent NDV and IBV. This study indicated that N-2-HACC-CMC could be used as an efficient adjuvant and delivery carrier for further development of mucosal vaccines and drugs and could have an immense application potential in medicine.     

鸽Ⅰ型副粘病毒的研究

本文探讨了鸽Ⅰ型副粘病毒(鸽PMV—1)人工感染病鸽的临床症状,鸽PMV—1的抗原性、生物和理化特性以及对鸡的致病性,并与鸡新城疫病毒(NDV),特别是速发型亲内脏性新城疫病毒株(VVNDV)进行了比较。此外,初步测试了几种常用的鸡新城疫疫苗对鸽抗NDV或抗鸽PMV—1的免疫效果;对广州、深圳地区的鸽子进行了鸽PMV—1感染的血清学调查。     

DNA-chitosan nanoparticles improve DNA vaccine-elicited immunity against Newcastle disease virus through shuttling chicken interleukin-2 gene

In this study, pCAGG-ChIL2 plasmid DNA containing the chicken interleukin-2 (ChIL-2) gene was used to prepare DNA-chitosan nanoparticles (CNPs). The CNPs prepared were spherical, with mean diameters between 100 and 200?nm, have a positive surface charge, and could protect DNA against DNase I degradation. The CNPs prepared were successfully used to transfect the Df-1 cell line with almost no cytotoxicity. CNPs prepared at an amino group to phosphate group ratio (N/P ratio) of 16 provided the highest transfection efficiency (1.1%) in medium with a pH of 6.5. When pCAGG-ChIL2 CNPs were administered to chickens simultaneously with a DNA vaccine against Newcastle disease virus (NDV), haemagglutination inhibition antibody titers and serum interferon-γ (IFN-γ) levels were significantly higher than in chickens immunised with the NDV DNA vaccine alone (p?相似文献   

DNA–chitosan nanoparticles improve DNA vaccine-elicited immunity against Newcastle disease virus through shuttling chicken interleukin-2 gene

In this study, pCAGG-ChIL2 plasmid DNA containing the chicken interleukin-2 (ChIL-2) gene was used to prepare DNA–chitosan nanoparticles (CNPs). The CNPs prepared were spherical, with mean diameters between 100 and 200?nm, have a positive surface charge, and could protect DNA against DNase I degradation. The CNPs prepared were successfully used to transfect the Df-1 cell line with almost no cytotoxicity. CNPs prepared at an amino group to phosphate group ratio (N/P ratio) of 16 provided the highest transfection efficiency (1.1%) in medium with a pH of 6.5. When pCAGG-ChIL2 CNPs were administered to chickens simultaneously with a DNA vaccine against Newcastle disease virus (NDV), haemagglutination inhibition antibody titers and serum interferon-γ (IFN-γ) levels were significantly higher than in chickens immunised with the NDV DNA vaccine alone (p?<?0.05). The results demonstrate that pCAGG-ChIL2 CNPs improve DNA vaccine-elicited immunity against NDV challenge.     

Virosome and ISCOM vaccines against Newcastle disease: preparation, characterization and immunogenicity

The purpose of this study was to prepare and characterize virosomes and ISCOMs containing envelope proteins of Newcastle disease virus (NDV) and to evaluate their immunogenicity in target animals (chickens). Virosomes were prepared by solubilization of virus with either Triton X-100 or octyl glucoside (OG) followed by detergent removal. Biochemical analysis revealed that these virosomes contained both the haemagglutinin-neuraminidase protein (HN) and the fusion protein (F), with preserved biological activity. Acidic environment triggered the fusion between virosomes and chicken erythrocyte ghosts. Formation of ISCOMs was achieved by solubilizing phospholipids, cholesterol, envelope protein antigen and Quil A in Triton X-100. The ISCOM particles were formed by removal of the detergent. In each formulation the relative HN content correlated with the capability to agglutinate red blood cells. The immunogenicity of these lipid-based subunit vaccines was determined in chickens after subcutaneous immunization. The relative HN content of the subunit vaccines correlated with the haemagglutination-inhibition (HI) antibody titres. Virosomes prepared with Triton X-100 and ISCOMs offered high clinical protection (> 80%) upon challenge with virulent NDV. Virosomes prepared with OG yielded lower clinical protection despite high HI antibody titres. Virosomes with reduced antigen density showed poor immunogenicity and protection. In conclusion, ND virosomes and ISCOMs were found to be immunogenic and provided good protection.     

Human effects of veterinary biological products

A 14-year-old male drank two glasses of milk from a gallon inoculated with 21 vials of live virus vaccine intended to immunize 1000 baby chicks against Newcastle Disease. The patient was managed with catharsis and careful observation, and remained completely asymptomatic in the following 28 days. Newcastle Disease virus (NDV) may cause paralytic neurotropic, viscerotropic, or pneumonotropic disease in chickens, but has limited effects on humans. Systemic exposure to this vaccine has not previously been reported in humans, although an aerosol NDV vaccine has caused self-limited conjunctivitis following ocular exposure. The human effects of veterinary biologicals remain poorly defined. The limited literature existing in regards to the human experience with common veterinary vaccines is reviewed.     

Naturally occurring mutations at residues 253 and 284 in VP2 contribute to the cell tropism and virulence of very virulent infectious bursal disease virus

Infectious bursal disease virus (IBDV) is responsible for the highly contagious infectious bursal disease in chickens. Previously, by blind passage, a vvIBDV Gx strain was attenuated to the Gt strain, and a strain CEF-9 with intermediate characters was obtained during attenuation. Since CEF-9 exhibited only two interesting amino acid mutations (Q253H and A284T) on loops P_DE and P_FG at the tip of VP2 spikes, we hypothesized that, either function separately or in combination, they define the cell tropism and virulence of vvIBDV. To test this hypothesis, Q253H and A284T were introduced individually or in combination into VP2 of the Gx or Gt strain to obtain six modified clones. Using reverse genetics, combined mutations of Q253H and A284T could adapt vvIBDV to non-permissive CEF cells (rGx-F9VP2) but any single mutation could not. In vivo, rGx-F9VP2 did not cause mortality while the Gx strain induced 66.7% mortality. Dual evidence from natural and rescued strains identified that the cell tropism of vvIBDV to CEF cells was determined by the combined VP2 mutations Q253H and A284T, but not by single mutation. The two residues were mainly responsible for the virulence of vvIBDV. These findings may be helpful in the design of new tailored IBDV vaccines.     

高滴度特异性卵黄抗体在天花病毒检测中的研究

天花是人类第一个消灭的疾病,但生物界中仍广泛存在天花类病毒。目前人类还缺乏对其有效、快捷的诊断、检测和治疗措施。在全球面临生物恐怖袭击的时代,这方面的研究工作尤显迫切。以3种灭活的天花病毒（vaccinia virus,calpox virus和cowpox virus）为免疫原,分别免疫注射3组实验蛋鸡,以改良的聚乙二醇法将相应特异性抗体从卵黄中提取。利用免疫荧光实验、抗体中和实验及免疫电镜等实验持续检测抗体滴度变化以及抗体交叉反应。最后,以特异性的卵黄抗体与磁珠（Dynabead）相结合,用于天花病毒的纯化,并在PCR实验中验证其检测效果。抗vaccinia virus抗体和抗calpox virus抗体在高度稀释的情况下（分别以1：10^6,1：10^5稀释）,仍在免疫荧光实验中呈阳性反应,且抗体的这种高表达水平在持续5次免疫注射下,能维持达10个月。抗体的中和能力和抗体与抗原反应的超微结构也在相应实验中观察到。抗体与病毒蛋白结合的特殊条带也在免疫印记实验中观察到。实验显示,不同天花病毒之间存在强而明显的交叉反应。最后,特异性的卵黄抗体能够与磁珠结合,并在模拟样本中用于天花病毒的PCR检测。达到纯化病毒、增强PCR实验敏感度的效果。该研究显示,抗天花病毒卵黄抗体能够应用于天花病毒的诊断,有望以此为基础,开发出快速、有效的天花病毒检测手段。     

麻疹腮腺炎减毒活疫苗生产用鸡胚的质量控制研究

目的  建立麻疹腮腺炎减毒活疫苗生产用鸡胚的质量控制方法。方法  运用酶联免疫吸附测定检测禽白血病病毒;运用双向免疫扩散试验检测马立克氏病病毒、网状内皮组织增生症病毒、禽流感病毒、传染性法氏囊病病毒、多杀性巴氏杆菌;运用血凝试验检测血吸附病毒;运用逆转录-聚合酶链反应检测新城疫病毒;依照《中华人民共和国药典》2010年版三部的要求进行无菌检查及支原体和外源因子检查。结果  采用上述方法未在无特定病原体鸡胚（鸡群代码：S、R、D、W、C、F、G、M、H）尿囊液、原代鸡胚成纤维细胞悬液、鸡胚成纤维细胞悬液培养物中检出禽白血病病毒、马立克氏病病毒、网状内皮组织增生症病毒、禽流感病毒、传染性法氏囊病病毒、多杀性巴氏杆菌、血吸附病毒和新城疫病毒,表明采用这些方法对麻疹腮腺炎减毒活疫苗生产用鸡胚进行质量控制是可行的。结论  建立了一套用于麻疹腮腺炎减毒活疫苗生产用鸡胚质量控制的快速、特异的检测方法。