豚鼠醛化红细胞的制备及其在新城疫和禽流感HA/HI试验中的应用

目前,在新城疫(ND)和高致病性禽流感(HPAI)的疫情免疫监测中,血凝/血凝抑制试验(HA/HI)依然是基层畜牧兽医部分最主要的检测方法,β-微量法以其操作简便、迅速、易判定、能准确表达结果等优点被广泛应用。在HA/HI试验中,需要使用1%鸡新鲜红细胞作指示剂。但鸡新鲜红细胞存在着以下几个缺点:一是健康非免疫公鸡较难获得,尤其是不同鸡或同一羽鸡不同时期采的红细胞,用于实验可能存在较大的差异;二是新鲜鸡红细胞质脆易碎,保存期短,需临用时现采现配;三是新鲜鸡红细胞易吸附免疫球蛋白等物质,洗涤不彻底易影响实验结果。由此导致了血凝/…     

鸡醛化红细胞在血凝和血凝抑制试验中的应用

应用鸡醛化红细胞 (FoRBC)代替新鲜配制红细胞 (FrRBC) ,进行血凝试验 (HA)和血凝抑制试验 (HI) ,以检测病毒滴度及其对应抗体效价。结果 :0 75 %FoRBC与 1 %FrRBC所测病毒HA和相应抗体HI效价一致。表明 0 75 %FoRBC可以替代 1 %FrRBC ,且具有不易破裂 ,保存期长 ,使用方便 ,无自凝 ,图象清晰、稳定 ,结果易判定等优点     

在鸡新城疫HA和HI试验中使用醛化红细胞的试验观察

长期以来,在鸡新城疫(ND)血凝(HA)和血凝抑制(HI)试验中均使用新鲜鸡红细胞,由于这种新鲜红细胞保存时间短,使用之前临时采血洗涤,费时又不方便;同时因不同批次或不同鸡只个体红细胞的差异而影响检测结果。而醛化红细胞具有保存时间长(一年左右)、不溶血、不易破碎和使用方便等优点。笔者在鸡新城疫 HA 和 HI 试验中使用醛化红细胞替代新鲜红细胞进行了试验观察,获得满意的效果,现报告如下。     

鸡醛化红细胞在鸽Ⅰ型副粘病毒HA和HI试验中的应用

在血凝（HA）和血凝抑制（HI）试验中，传统的方法采用新鲜红细胞。由于新鲜红细胞容易破碎，保存时间短，每次试验需临时准备，使用很不方便。本文采用醛化红细胞进行鸽1型副粘病毒的HA和HI试验，并与新鲜红细胞进行对照，结果表明，两种红细胞在鸽1型副粘病毒的HA和HI试验中的结果基本一致，而使用醛化红细胞更为便利。现将结果报道如下。1材料与方法l．l鸽1型副粘病毒（鸽A／PMV－1）；鸽A”MV－IZH株由本室鉴定保存，在9－11日龄鸡胚培养后收获尿囊液无菌检查后置4C保存备用。用于HI试验的病毒经0．2％甲醛37C灭活24小时后置…     

豚鼠醛化红细胞在人流感病毒血凝活性测定中的应用

在流感疫情监测中,血凝试验是最常用的检测方法.针对鸡红细胞不能凝集流感病毒"O"相毒株,新鲜红细胞需现采现配、麻烦、费时,易溶血、不易保存等问题,本试验对豚鼠醛化红细胞制备方法的筛选和制备条件的优化进行研究,并以新鲜豚鼠红细胞为对照,用于流感病毒不同型别血凝试验.结果表明,戊二醛固定法制备的豚鼠醛化红细胞不溶血、结果稳定性好、保存期长、血凝活性与新鲜豚鼠红细胞基本一致,可以替代新鲜豚鼠红细胞用于血凝试验.     

醛化鸡红细胞应用于鸡新城疫的诊断和抗体检测

将新鲜鸡红细胞经戊二醛处理后,制成醛化鸡红细胞。该醛化鸡红细胞具有特异性,不发生自凝,保存期可达半年以上,和阴性尿囊液不发生凝集,与新工细胞同时用于血球凝集和血球凝集抑制试验,对疑似鸡新城疫病料和待检测血清进行检测,结果在ＨＡ试验中,新鲜鸡红细胞比醛化鸡红细胞高１－３个滴度,而在ＨＩ试验中,两种红细胞所得结果基本一致,有的则醛化对外开放红细胞比新鲜鸡红细胞高１－２个滴度。由此说明,醛化鸡红细胞完全     

对血凝试验用鸡红细胞保存条件的探讨

对血凝试验所用的鸡红细胞的保存条件进行了比较。在新鲜红细胞液中加入甘油或尿素可以延缓细胞破裂溶血。鸡红细胞经戊二醛处理后，于4℃保存9个月，其凝集特性未发生变化，同时在醛化红细胞中加入甘油更能稳定血凝活性。在4℃、室温和37℃条件下将制备的新鲜红细胞和醛化红细胞用作血凝试验的指示剂，不影响新城疫病毒抗原血凝滴度。结果表明，所制备的醛化鸡红细胞的甘油悬液可以替代新鲜红细胞用于临床实践中新城疫的血凝和血凝抑制试验测定。     

Proteomic analysis of chicken peripheral blood mononuclear cells after infection by Newcastle disease virus

Characteristic clinical manifestations of Newcastle disease include leukopenia and immunosuppression. Peripheral blood mononuclear cells (PBMCs) are the main targets of Newcastle disease virus (NDV) infection. To survey changes in proteomic expression in chicken PBMCs following NDV infection, PBMC proteins from 30 chickens were separated using two-dimensional electrophoresis (2-DE) and subjected to mass spectrometry analysis. Quantitative intensity analysis showed that the expression of 78 proteins increased more than two-fold. Thirty-five proteins exhibited consistent changes in expression and 13 were identified as unique proteins by matrix assisted laser desorption ionization-time of flight mass spectrometer/mass spectrometer including three that were down-regulated and 10 that were up-regulated. These proteins were sorted into five groups based on function: macromolecular biosynthesis, cytoskeleton organization, metabolism, stress responses, and signal transduction. Furthermore, Western blot analysis confirmed the down-regulation of integrin-linked kinase expression and up-regulation of lamin A production. These data provide insight into the in vivo response of target cells to NDV infection at the molecular level. Additionally, results from this study have helped elucidate the molecular pathogenesis of NDV and may facilitate the development of new antiviral therapies as well as innovative diagnostic methods.     

陕北地区鸡新城疫免疫监测及分析

以神木县为例,介绍了陕北地区鸡新城疫的防疫情况,揭示新城疫防疫的薄弱环节。本研究从神木县多家种鸡场、商品蛋鸡场及活禽交易市场采集血清样本346份,用血凝抑制试验测定新城疫抗体滴度,结果显示商品蛋鸡场免疫效果较差,合格率只有69.6%;同时发现种鸡场也存在免疫问题。在今后新城疫防控过程应与重视。     

Studies on the Pathogenicity of Newcastle Disease Virus Isolates in Guinea Fowl

Newcastle disease viruses isolated from chickens and guinea fowl were characterized as viscerotropic, velogenic strains on the basis of their mean death time, intracerebral pathogenicity index, intravenous pathogenicity index and cloacal and conjunctival mean death time. The pathogenesis of the disease caused by both the strains was studied in 4-week-old guinea fowl. Both strains had an incubation period of 5 days and the birds showed dullness, depression, anorexia, diarrhoea and paralysis of the legs. They also exhibited nervous signs such as incoordination, muscle tremors and trembling of the neck at the advanced stage of the disease. Mortality reached 52% in the group infected with the chicken isolate but it was only 8% in the birds infected with the guinea fowl isolate. No specific changes were observed at post-mortem examination except haemorrhages at the tip of the glands of the proventriculus and in the caecal tonsil. Changes in the lymphoid organs and brain were always present in both the groups. Despite the low mortality, the guinea fowl isolated had multiplied in various organs in the birds. In both groups, the frequency of virus isolation increased from 5 to 10 days post infection.     

新城疫病毒复制对其溶瘤作用影响的初步分析

本研究旨在明确新城疫病毒（NDV）溶瘤作用是否依赖其复制水平,并改进NDV溶瘤机制研究模型。以NDV疫苗株LaSota感染人肿瘤细胞系HCT116、A549和人非肿瘤细胞系HEK293T为研究模型;RT-qPCR检测NDV在各细胞系的基因组复制水平;Western blot检测NDV在各细胞系的蛋白表达水平;利用流式细胞术检测NDV感染对各细胞系的细胞周期和细胞凋亡的影响。结果表明,NDV在三个细胞系内的基因组复制水平和蛋白表达水平没有显著差异,但对三个细胞系的溶瘤作用存在明显差异;进一步流式细胞术检测发现,NDV感染能诱发HEK293T和HCT116的细胞周期发生G1期停滞,但对A549的细胞周期没有明显影响;此外,NDV感染24 h后,A549细胞以早期凋亡为主,而HCT116细胞以坏死/晚期细胞凋亡为主。NDV的溶瘤作用并不依赖于其复制水平,而且NDV对不同类型肿瘤细胞的溶瘤作用存在不同机制。     

一株鸡新城疫病毒的分离鉴定及其油乳剂灭活苗免疫效果评价

从河南某种鸡场发病鸡群中分离出1株毒株,经HA、HI和动物回归试验,确定分离的病毒为NDV,命名为HNYD株。经过测定,HNYD株鸡胚半数致死量（ELD50）为10^90/0．1mL;鸡胚最小致死量的平均致死时间（MDT）为53．8h;对1日龄SPF雏鸡脑内致病指数（ICPI）为1．86,提示HNYD株为强毒力株。血清交叉反应试验显示HNYD株和La Sota株存在一定的抗原性差异。用HNYD株油乳剂灭活苗对HNYD株和La Sota株的保护率分别为90％和100％,明显优于La Sota株油乳剂灭活苗,提示HNYD株制备的疫苗对流行毒株和经典毒株的攻击均能很好的保护。     

Dot—ELISA间接法检测鸡新城疫病毒（NDV）的研究

常规方法分离纯化鸡ＩｇＧ,免疫羊制备羊抗鸡ＩｇＧ二抗,以辣根过氧化物酶（ＨＲＰ）标记制备酶标记物的工作浓度为１：１０００;选用硝酸纤维素（ＮＣ）膜作固相载体,建立检测ＮＤＶ抗原的斑点间接酶联免疫吸附试验诊断方法。检测ＮＤＶ鸡胚毒４０份,阳性检出率１００％;人工感染非免疫鸡的气管组织、肺组织各４７份,检出率为９３．６％;人工感染ＳＰＦ鸡气管组织、肺组织各４０份,检出率１００％。与传染性支气管炎病毒（ＩＢＶ）、减蛋综合症病毒（ＥＤＳＶ）、传染性法氏囊病病毒（ＩＢＤＶ）无交叉反应。试验显示该方法简便、特异、快速、结果直观,便于生产应用     

Assessment of the red blood cell proteome in a dog with unexplained hemolytic anemia

A 7‐year‐old female neutered Jack Russell Terrier was presented to Langford Vets, the University of Bristol, with a history of chronic intermittent lethargy. Investigations and clinical course were compatible with hereditary hemolysis due to a red blood cell membrane defect. Proteomics was used to explore protein alterations in the presence of a hypothesized red blood cell membrane protein deficiency. Proteomic analysis revealed downregulation of the band 3, and alpha‐ and beta‐adducin proteins, and alterations in the red blood cell proteome consistent with previous reports of changes due to the presence of reticulocytosis and ongoing hemolysis. The spectrum of protein alterations identified in the affected dog may be homologous to a band 3 protein deficiency secondary to hereditary spherocytosis, as described in people.     

不同佐剂新城疫疫苗对鸡体液免疫的影响

佐剂是疫苗研究的重要组成部分.选择合适的佐剂不仅可以增强疫苗的免疫效果,而且可以提高机体的抵抗力.不同佐剂对体液免疫会产生不同的作用.本研究选用白油佐剂、弗氏佐剂和蜂胶佐荆与新城疫病毒抗原混合制成疫苗,免疫雏鸡,探讨不同佐剂对鸡体液免疫的影响.结果表明,三种佐剂疫苗均有保护力,试验鸡饲养至61日龄均未发生新城疫,其中弗氏佐剂的效果最好,蜂胶佐剂的效果次之,油佐剂组检测到雏鸡61日龄时仍未出现抗体高峰值.     

复方中药体外抗新城疫病毒活性研究

为了筛选出高效的抗新城疫病毒中药,采用鸡胚接种试验,对8种经典清热解毒复方中药的抗新城疫病毒效果进行了测定。结果表明,荆防败毒散、银翘散、白头翁汤、双黄连口服液在体外对新城疫病毒具有较好的抑制作用。