抗猪传染性胃肠炎病毒与猪流行性腹泻病毒卵黄抗体制备及其临床应用研究

本研究旨在制备猪传染性胃肠炎病毒和猪流行性腹泻病毒高免卵黄抗体,研究其治疗效果。以猪传染性胃肠炎和猪流行性腹泻二联灭活苗免疫产蛋鸡,琼脂扩散方法检测抗体效价达到1∶64时,收集卵黄,采用氯仿抽提和硫酸铵盐析法纯化卵黄抗体,进行微生物学检测、安全性试验,通过人工感染治疗试验和临床应用,观察其治疗效果。结果人工感染治愈率为100%,临床应用治愈率为88.0%,表明制备的卵黄抗体对猪传染性胃肠炎和流行性腹泻具有显著的治疗效果。     

二种方法检测猪传染性胃肠炎病毒抗体的比较

分别用血清中和试验和单克隆抗体阻断酶联免疫吸附试验对８１头美国进口猪轿清作猪传染性胃肠炎抗体检测。ＳＮ试验同７份阳性血清，检出率为８．６４％，Ｂ－ＥＬＩＳＡ试验检出７份ＰＲＣＶ抗体阳性血清，检出率为８．６４％，无ＴＧＥ阳性。ＳＮ试验检出７份ＴＧＥ抗体阳性血清与Ｂ－ＥＬＩＳＡ试验检出的７份ＰＲＣＶ抗体阳性血清完全重合。结果证明，应用单克隆抗体进行的Ｂ－ＥＬＩＳＡ可鉴别诊断ＴＧＥ与ＰＲＣＶ感染，优于Ｓ     

SPA-ELISA监测猪传染性胃肠炎病毒血清抗体水平的研究

建立了SPA—ELISA监测猪传染性胃肠炎病毒(TGEV)血清抗体水平的方法，与免疫荧光试验(IFA)符合率高达90．9％。试验表明：病毒抗原最适包被浓度为4μg／ml；血清工作浓度为1：200；最佳包被液采用0．05m pH9．6的碳酸缓冲液；封闭液选用2％的脱脂奶；并确定了抗原抗体最佳反应时间和最适反应温度及底物溶液的显色时间。特异性试验表明所组装的SAP—ELISA试剂盒特异性较好，可用于TGE的流行病学调查。     

猪传染性胃肠炎病毒核酸免疫昆明鼠的抗体应答

以猪传染性胃肠炎病毒(TGEV)TH98株N基因的重组质粒pHN为模板，Li-15和Li-2为上、下游引物扩增出TGEVTH98／N基因。将N基因与真核表达载体pcDNA3．1( )分别用BamHⅠ、XhoⅠ双酶切后连接，构建了重组真核表达载体pcDNA—N。将昆明鼠随机分为2组，分别肌肉注射pcDNA—N和pcDNA3．1( )，每只鼠100μg，共免疫3次，每次间隔2周。首次免疫后第0、14、21、28、35、39、47和54d采血分离血清，采用ELISA检测抗体动态变化。注射pcDNA—N组小鼠在首次免疫后第39d检测到针对TGEVN蛋白的阳性抗体。     

猪传染性胃肠炎的防治

猪传染性胃肠炎(TGE)是由猪传染性胃肠炎病毒(TGEV)引起的一种高度传染性病毒性肠道疾病,以导致2周龄以下仔猪呕吐、严重腹泻和高病死率(通常为100%)为主要特征。养猪场一旦发生该病将造成重大损失。该文对猪传染性胃肠炎的防治进行综述,供同行参考。     

猪传染性胃肠炎研究进展

猪传染性胃肠炎病是由猪传染性胃肠炎病毒引起的,是一种急性、高接触性肠道传染病。该病的爆发具有季节性,对1周龄以内新生仔猪危害尤为严重,感染之后能使仔猪发生的呕吐、腹泻和脱水的症状,严重的死亡率可达100％,对新生仔猪的危害尤为严重,并对当前养猪生产造成了特大的经济损失。从猪胃肠炎的流行病学、病毒特性、疾病检测及猪胃肠炎疫苗研究进展进行了综述,并对猪胃肠炎病毒的研究进行展望。     

猪传染性胃肠炎的综合防治

文章介绍了猪传染性胃肠炎发病特点及临床症状,并提出了综合治疗措施。     

猪传染性胃肠炎的防治

猪传染性胃肠炎(TGE)是由猪传染性胃肠炎病毒(TGEV)引起的猪的一种急性、高度接触性传染病，主要临床症状为呕吐、严重腹泻和脱水。不同年龄和品种的猪对本病均易感，其中2周龄以内的仔猪致死率可达100％，通常死于脱水和电解质代谢紊乱，给养猪业造成了极大的危害。现将防治方法介绍如下，供大家参考。     

猪传染性胃肠炎病毒分子生物学研究进展

本文对猪传染性胃肠炎病毒有关分子生物学方面的最新研究资料进行了概述。ＴＧＥＶ基因组约２９大小,其５‘端是基因１,余下３’端约８．３ｋｂ有６人基因,共有７个亚基因组ｍＲＮＡ,其转录,翻译过程受到病毒的自身的严格调控。基因１编码病毒的主要功能蛋白－聚合酶类有木瓜蛋白酶,类３Ｃ蛋白酶,类生长因子／受体区,聚合酶,金属离子结合区和解旋酶等功能活性区,它们对该酶活性进行调控。     

猪传染性胃肠炎病毒单克隆抗体的制备与鉴定

以纯化的猪传染性胃肠炎病毒(TGEV)作为抗原免疫BALB/c小鼠,经3次免疫后,通过聚乙二醇方法进行融合,利用有限稀释法在HAT培养基上筛选杂交瘤细胞,共获得12株既能稳定生长并可以分泌特异性抗TGEV的单克隆抗体的杂交瘤细胞系,分别命名为8D2、4H4、5D6、7H10、4C3、9G1、3A2、8G1、5G6、2D7、4D6、6B10。间接ELISA、Western blot、间接免疫荧光结果表明,获得的12株单抗能特异性识别TGEV,通过间接ELISA做病原检测,显示12株单抗与猪流行性腹泻病毒(PEDV)和猪轮状病毒(PrV)不发生交叉反应。经抗体亚类鉴定,该12株单克隆抗体均为IgG2b。12株抗TGEV的单抗制备成功,为猪传染性胃肠炎病原特性研究和病原快速检测提供了物质基础。     

Detection of antibody against transmissible gastroenteritis virus of pigs by indirect immunoperoxidase antibody test

Immunoperoxidase intibody (IPA) test was developed for detecting antibody against transmissible gastroenteritis (TGE) virus of pigs. The IPA antibody titers in sera collected in the field from 82 pigs were approximately seven times higher than those obtained in a serum-neutralization test. The correlation between the TGE antibody concentrations in the IPA and serum neutralization tests was positive (r = +0.74). The IPA tests appears to have the potential for routine laboratory use for serologic diagnosis of TGE.     

Molecular biology of transmissible gastroenteritis virus

The causative agent (TGEV) of porcine transmissible gastroenteritis belongs to the Coronaviridae, a family of enveloped viruses with a positive, single-stranded RNA genome. Important progress has recently been made concerning the molecular biology of TGEV. The research work of our group has been focused on two main aspects: genome structure and functional domains of the envelope proteins. TGEV genomic RNA is organised into seven regions. The sequence of six of them, i.e. the 3' most 8300 nucleotides, has been established from cDNA clones. Three genes encoding the structural proteins, the peplomer protein E2, the transmembrane protein E1 and the nucleoprotein, have been identified. Additional open reading frames allowed for the prediction of four non-structural polypeptides, the role of which remains to be discovered. The remaining part of the genome (estimated length 20 kb) is thought to encode the polymerase. Expression of TGEV genes involves the production of six subgenomic mRNAs, which together with the virion RNA, form a 3' terminal nested set. The peplomer glycoprotein E2 (220 kDa) is 1431 residues long and highly glycosylated. Several domains were identified, including a C-terminal anchoring region and at least four major antigenic sites, which cluster in the amino half part of the molecule. Two sites containing most of the critical neutralisation determinants are highly conserved among TGEV strains. The glycoprotein E1 (29kDa) is mostly embedded in the membrane and plays a crucial role in the virion architecture. However, a short N-terminal domain protruding out of the particle mediates complement-dependent neutralisation, and induces alpha interferon synthesis, likely through a direct interaction with the lymphocyte membrane.     

猪传染性胃肠炎病毒(TGEV)研究进展

猪传染性胃肠炎 (TGE)是由猪传染性胃肠炎病毒 (TGEV)引起的一种以严重腹泻、呕吐和脱水为临床特征的高度接触性传染病。 TGE首次由Doyle和 Hutchings1 946年在美国报道 ,日本 (1 95 6年 )和英国 (1 95 7年 )先后报道该病 ,这之后欧洲、北美、亚洲等多个国家相继报道发生了 TGE,现已成为一种世界性猪的疾病。我国从 6 0年代起就有TGE的报道 ,近些年来有进一步流行的趋势 ,尤其是冬季和早春寒冷季节常呈地方性暴发流行 ,给养猪业造成极大危害。在 1 984年和 1 986年间 ,欧洲北美等地又报道了一种特别的 TGEV自然缺失变异株 -猪呼吸…     

Inhibition of virus attachment to CPK cells by monoclonal antibody to transmissible gastroenteritis virus.

Five MAbs, which recognized E2 glycoprotein of TGE virus TO-163 and showed neutralizing activity, were examined to see if they inhibit virus attachment to the susceptible cells (CPK cells). Only one (160/4) MAb blocked the virus attachment to the cells, indicating that the inhibition of virus attachment is one of important mechanisms of neutralizing by antibody.     

Enzyme-linked immunosorbent assay for detection of transmissible gastroenteritis virus antibody in swine sera

An enzyme-linked immunosorbent assay (ELISA) was developed for detection and quantification of serum antibodies to transmissible gastroenteritis virus (TGEV) in swine. Sera from pigs inoculated with cell culture-origin TGEV or gut-origin TGEV were tested for anti-TGEV antibody by ELISA and by serum virus-neutralization test (NT). The ELISA detected antibody 3 days (av) sooner than did the NT when sera from pigs inoculated with cell culture-origin TGEV were tested and 1 day sooner than did the NT when sera from pigs inoculated with gut-origin TGEV were tested. The ELISA appeared to be more sensitive than the NT, since ELISA was more responsive to low-level antibody and ELISA titers exceeded NT titers.     

Methanol precipitation of transmissible gastroenteritis virus.

Methanol precipitation of transmissible gastroenteritis virus was tested at Ph 4.0, 5.0, 6.0, and 7.0 and at methanol concentrations of 15%, 25%, and 30%. Supernatant and precipitate fractions were tested for complement-fixing and agar-diffusion soluble antigens and plaque-forming units, and were examined by electron microscopy. Virus could be obtained free of detectable agar-diffusion antigens and most of the complement-fixing antigens. Most of the virions were without peplomers after methanol treatment but they retained infectivity.     

猪传染性胃肠炎病毒的诊断研究进展

猪传染性胃肠炎（Swine transmissble gastroenteritis,TGE）是由猪传染性胃肠炎病毒（Swine transmissble gastroenteritis virus,TGEV）引起的主要以2周龄以内的仔猪发生呕吐、严重腹泻和脱水为特征的一种高度接触性病毒传染病。1933年,美国的依利诺斯州就有本病的报道,