共查询到19条相似文献，搜索用时 343 毫秒
1.
为快速有效测定猪瘟疫苗病毒含量,建立了猪瘟兔化弱毒(HCLV)间接免疫荧光试验(IFA)方法,并与兔体定型热试验方法进行了初步比较。试验结果表明,以冷甲醇固定ST细胞,将所选猪瘟活疫苗检验用阳性血清1:40倍稀释、荧光二抗1:32倍稀释时,IFA检测HCLV感染的ST细胞清晰、特异,检测猪伪狂犬病毒、猪流行性腹泻病毒、猪传染性胃肠炎病毒感染细胞阴性。对不同猪瘟疫苗半成品的检测结果显示,IFA测定半数组织感染量(TCID50)与兔体定型热试验测定兔体感染量(RID)具有较好的相关性,其拟合曲线为RID/m L=2.783TCID50/m L+110107.213。 相似文献
2.
本研究使用间接免疫荧光方法(IFA)和过氧化物酶单层试验(IPMA)测定猪瘟兔化弱毒的组织半数感染量(TCID50)。优化了两种方法的一抗使用浓度;筛选了IPMA方法中敏感、易辨识的酶作用底物。优化后的IFA和IPMA方法测定猪瘟兔化弱毒疫苗半成品和成品的TCID50,可以作为兔体感染量(RID)和猪体半数保护量(PD50)效力检验标准的补充,应用到猪瘟兔化弱毒活疫苗生产中,加强质检、质控力度。 相似文献
3.
在做荧光抗体检测病毒中和试验时,用不同毒株检测同一份血清样品经常出现不一样的效果。通过开展猪瘟病毒(兔化弱毒株)和猪瘟病毒(法国Thiveral株)在ST细胞上的增殖研究,掌握猪瘟活疫苗收获抗原的最佳时间,选择更好的毒株进行荧光抗体检测病毒中和试验。结果表明:猪瘟病毒(兔化弱毒株)增值效果差,猪瘟病毒(法国Thiveral株)繁殖速度很快,在接种后30 h就能达到10~(5.4)TCID_(50)/mL,55 h达到最高峰10~(6.0)TCID_(50)/mL,72 h后呈平稳状态。 相似文献
4.
1猪瘟活疫苗（Ⅰ）
1.1名称：通用名,猪瘟活疫苗（Ⅰ）
1.2简介：本品系用猪瘟兔化弱毒C株接种乳兔,收获感染乳兔实质脏器制成乳剂,加适宜稳定剂,经冷冻真空干燥制成,因此又称猪瘟兔化弱毒乳兔组织疫苗。每头份疫苗所含病毒稀释150倍后能引起兔体的热反应。 相似文献
5.
RT-PCR法快速检测与鉴别诊断猪瘟病毒野毒株和三个兔化弱毒疫苗株的研究 总被引：1，自引：0，他引：1
基于猪瘟病毒基因组中富含T的插入序列建立起的简捷一步法RT-PCR技术,用于同步检测与鉴别野毒株和疫苗株。依据猪瘟病毒兔化弱毒疫苗株3′NTR独立存在富含T的插入序列,设计特异性的扩增引物,使用简捷一步法RT-PCR或巢式PCR之后用琼脂糖凝胶电泳或多毛细管电泳方法进行分析,能检测到并且准确鉴别在临床标本中的古典猪瘟病毒野毒株和兔化猪瘟疫苗株。这些检测技术至少可用于3个兔化弱毒疫苗株LPC、HCLV和C-株的检测。野毒株RT-PCR检测限量是6.3TCID50/mL,巢式PCR检测限量是0.63TCID50/mL。在前一次研究中,在猪瘟兔化弱毒疫苗株的基因组的3′NTR发现有12～13nt的富含T的插入序列;本次研究中,在LPC/PRK和LPC/TS兔化疫苗株的基因组中发现2个长为42和36nt富含T的插入序列。这些长为12、36、42nt富含T的插入序列增加PCR片段的大小,有利于遗传标记对猪瘟病毒野毒型和不同兔化疫苗株间的快速检测和鉴别。 相似文献
6.
单克隆抗体间接免疫荧光检测猪瘟病毒方法的建立 总被引：1，自引：0，他引：1
以作者制备的抗猪瘟病毒单克隆抗体(anti-CSFV McAb)为一抗、荧光素标记羊抗小鼠IgG为二抗,通过反应条件的优化,建立检测猪瘟病毒抗原的间接免疫荧光(IFA)检测方法。确定IFA最佳工作条件:CSFV最佳接种浓度和培养条件为CSFV 10-3倍稀释后接种PK15细胞,37℃5%CO2恒温箱中培养36 h;McAb最适工作浓度为1∶1 000倍稀释;荧光素标记的羊抗小鼠IgG荧光抗体的最适工作浓度为1∶50倍稀释。特异性试验表明,用建立的IFA检测方法检测CSFV感染PK15细胞为阳性,而检测伪狂犬病病毒(PRV)、猪细小病毒(PPV)、猪2型圆环病毒(PCV-2)感染PK15细胞均为阴性。结果表明,建立的检测细胞培养中CSFV抗原的IFA检测方法具有敏感特异、简便快速等优点,可用于CSFV感染的实验室诊断及CSFV在感染细胞中的定位和动态分布研究。 相似文献
7.
猪瘟病毒猪细小病毒猪繁殖与呼吸综合征病毒猪伪狂犬病病毒多重PCR方法的建立以及初步应用 总被引：9，自引：0，他引：9
猪瘟病毒、猪细小病毒、猪繁殖与呼吸综合征病毒及猪伪狂犬病病毒均能导致猪繁殖障碍,对养猪生产影响很大。本研究通过设计4对针对这4种病毒的特异引物建立了多重PCR方法,分别对其最佳反应条件、特异性及敏感性进行了测定,结果表明:其敏感性可达到CSFV 10 TCID50,PPV 10TCID50,PRRSV 1 TCID50,PRV 100 TCID50 CSFV 10TCID50,PPV 10TCID50,PRRSV 1 TCID50,PRV 100 TCID50。同时具有较好的特异性,对猪瘟病毒石门株、猪瘟病毒兔化弱毒株、PRV闽南A株、PPV弱毒疫苗株、PRRSV KY 35株及PRRSV B13株6个毒株均能扩增出相应的片段,而BVDV、BDV均未扩增出相应的片段。本方法的建立对于这4种病毒病的早期快速检测具有十分重要的意义。 相似文献
8.
为建立一种能够区分猪瘟病毒(CSFV)强毒与弱毒疫苗C株的SYBR Green Ⅰ荧光定量RT-PCR结合熔解曲线分析方法,本研究对GenBank中登录的25株CSFV强毒株和兔化弱毒疫苗C株全基因组序列进行比较分析,设计一对共用下游引物以及分别针对CSFV强毒株与弱毒疫苗的特异性上游引物,其Tm值分别为84.5±0.5℃和88.5±0.5℃,熔解曲线分析显示为单特异峰.检测结果显示本实验建立的鉴别CSFV强毒感染与弱毒疫苗的荧光定量RT-PCR结合熔解曲线分析方法特异性强,对其他相关病毒无特异性扩增;敏感性高,最低检出量为5×RID50的细胞疫苗基因组拷贝;重复性好;并且扩增效率高、线性范围广、检测时间短,可对免疫猪群中CSFV强毒感染做出快速准确的鉴别检测,为有效防制猪瘟提供依据. 相似文献
9.
异源细胞培养猪瘟兔化弱毒活疫苗的保存期观察以体强（广东生物药厂广州，５１０６３０）在多年进行猪瘟细胞活疫苗的检验工作中，在研制羊肾细胞活疫苗作的－１５℃、８－１５℃保存期的测定中，发现猪瘟兔化弱毒株在异源细胞活疫苗中经较长时间的保存后毒价没有降低。在... 相似文献
10.
试验采用IFA法对2批随机抽样的PRRS活疫苗(每批4瓶)、阳性对照(CSFV)、阴性对照(未被CSFV污染的猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV))和特异性试验检测病毒(猪圆环病毒2型(porcine circo virus 2 type,PCV-2))接种于生长良好的PK-15细胞进行间接免疫荧光染色检测CSFV,以期建立快速、准确的间接免疫荧光法(indirect immunofluorescence assay,IFA)调查猪蓝耳病活疫苗中猪瘟病毒(classical swine fever virus,CSFV)的污染情况;在不同时间、相同试验条件下,相同批次的疫苗被重复检测了3次。试验结果表明:阳性对照接种的PK-15细胞出现特异性黄绿色荧光,阴性对照、PCV-2和待检样品接种的PK-15细胞均未出现特异性黄绿色荧光,即待检2批PRRS活疫苗未被CSFV污染;通过3次重复操作,最终的检测结果一致,说明IFA检测猪瘟病毒包涵体的特异性强、敏感性高。 相似文献
11.
A rapid biological assay based on incubation time has been developed for determination of the potency of Newcastle disease
virus strain I-2 vaccine. It is based on the observation that the interval between inoculation and the first detection of
haemagglutinin (HA) depends on the titre of the vaccine inoculated. Chicken embryonated eggs were inoculated with different
titres (109, 106 and 103 EID50/0.1 ml) of vaccine and incubated for 24 h. At hourly intervals, 5 eggs from each vaccine titre were tested for the presence
of HA. The results showed that the HA activity was detected from 5, 11 and 15 h after inoculation with vaccine doses of 109, 106 and 103 EID50, respectively. On the basis of these results it is suggested that if there is no HA detected from 5 to 11 h after inoculation
of eggs with the vaccine virus, the vaccine should not be used to vaccinate chickens as it might have an infectivity titre
of less than 106 EID50/0.1 ml, which is equivalent to the recommended single chicken dose. It is concluded that measuring the time between inoculation
of the vaccine virus and the onset of HA activity might provide an estimate of the titre of the vaccine within 24 h. 相似文献
12.
为了研究B亚群禽白血病病毒(ALV-B)的半数细胞培养物感染量(TCID50)与p27抗原的酶联免疫吸附试验(ELISA)的S/P值之间的相关性,本试验将ALV-B SDAU09C2 株接种鸡胚成纤维细胞(CEF细胞),换维持液后连续10 d取样,检测10 d的TCID50值与p27抗原的S/P值之间的相关性;同时,将该毒株在DF-1细胞系上传代至20代,取其中的第1、5、10、15和20代分别进行TCID50滴度的测定和p27抗原检测。结果表明,在CEF细胞上接种的ALV-B SDAU09C2株连续10 d的TCID50值与p27抗原之间存在显著的正相关(r=0.94002;P<0.0001);在DF-1细胞系上传的不同代数之间也呈显著正相关(r=0.96449;P=0.0080)。由此可推测ALV-B的TCID50与p27抗原呈显著正相关,可以用ELISA法测得的p27抗原的S/P值来估测病毒的TCID50值。 相似文献
13.
Yeast-expressed classical swine fever virus glycoprotein E2 induces a protective immune response 总被引：4，自引：0，他引：4
Guang-Jan Lin Ting-Yu Liu Yu-Yao Tseng Zeng-Weng Chen Chia-Chin You Shih-Ling Hsuan Maw-Sheng Chien Chienjin Huang 《Veterinary microbiology》2009,139(3-4):369-374
Classical swine fever (CSF) is an economically important swine disease worldwide. The glycoprotein E2 of classical swine fever virus (CSFV) is a viral antigen that can induce a protective immune response against CSF. A recombinant E2 protein was constructed using the yeast Pichia pastoris expression system and evaluated for its vaccine efficacy. The yeast-expressed E2 (yE2) was shown to have N-linked glycosylation and to form homodimer molecules. Four 6-week-old specified-pathogen-free (SPF) piglets were intramuscularly immunized with yE2 twice at 3-week intervals. All yE2-vaccinated pigs could mount an anamnestic response after booster vaccination with neutralizing antibody titers ranging from 1:96 to 1:768. Neutralizing antibody titers at 10 weeks post booster vaccination ranged from 1:16 to 1:64. At this time, the pigs were subjected to challenge infection with a dose of 1 × 105 TCID50 (50% tissue culture infective dose) virulent CSFV strain. At 1 week post challenge infection, all of the yE2-immunized pigs were alive and without symptoms or signs of CSF. Neutralizing antibody titers at this time ranged from 1:4,800 to 1:12,800 and even to 1:51,200 one week later. In contrast, the control pigs continuously exhibited signs of CSF and had to be euthanized because of severe clinical symptoms at 6 days post challenge infection. All of the yE2-vaccinated pigs were Erns antibody negative and had seroconverted against Erns by post challenge day 11, suggesting that yE2 is a potential DIVA (differentiating infected from vaccinated animals) vaccine. The yeast-expressed E2 protein retains correct immunogenicity and is able to induce a protective immune response against CSFV infection. 相似文献
14.
15.
本实验室对从疑似犬细小病毒感染的发病犬分离的病毒采用同步培养法接种猫肾细胞（CRFK）增殖,通过PCR试验、IFA试验和VP2基因测序分析等方法进行鉴定并分型,获得一株犬细小病毒强毒株,命名为CPV-DD株。对分离病毒进行PCR扩增,可扩增出特异性DNA片段（1 163 bp）;盲传至第6代时,病毒液的HA效价为1∶1... 相似文献
16.
SUMMARY Experiments were conducted with vaccines containing the V4 strain of Newcastle disease virus (NDV). Both living aqueous vaccines and vaccines consisting of virus incorporated in an oil emulsion were used. The calculated dose of virus contained in the oil emulsion vaccine was 108,7 50% embryo infectious doses (EID50) per bird dose. Haemagglutinin inhibition (HI) antibody levels of 8 are presumed protective. One-day-old chicks with low levels of maternal antibody were vaccinated intraocularly with 106,3EID50 of live vaccine, and concurrently with oil emulsion vaccine. Presumed protective levels of antibody were present at two weeks post vaccination and were maintained for at least seven weeks longer. When adult birds 15 weeks old with no previous exposure to NDV were vaccinated intraocularly with 106,7EID50 per bird, protective levels of antibody were produced within a week. Unvaccinated birds put in contact with the vaccinated birds produced similar antibody levels within 14 days. Revaccination with oil emulsion vaccine after antibody levels had fallen resulted in a rapid response with high levels of antibody. When antibody-free adult commercial birds with an unknown history of exposure to NDV were vaccinated intramuscularly with oil emulsion vaccine, high antibody levels were produced for at least 21 weeks. Concurrent intraocular inoculation with 107,0EID50 live virus did not enhance the response. Natural infection of unvaccinated birds occurred during the experiment. This was detected by the presence of HI antibody levels of short duration. When antibody-free commercial birds were inoculated intramuscularly with oil emulsion vaccine containing 106,0, 107,0, or 108,0EID50 per bird dose, 100% of birds inoculated with the highest dose produced presumed protective levels of antibody within two weeks, as compared with a 5-week delay when using the 107,0EID50 per bird dose. 相似文献
17.
Blacksell SD Khounsy S Van Aken D Gleeson LJ Westbury HA 《Tropical animal health and production》2006,38(6):467-474
This study investigated the comparative susceptibility of indigenous Moo Laat and improved Large White/Landrace pig breeds to infection with classical swine fever virus (CSFV) under controlled conditions
in the Lao People’s Democratic Republic (Lao PDR). The Moo Laat (ML) and Large White/Landrace crossbreed (LWC) pigs were inoculated with a standard challenge strain designated Lao/Kham225 (infectivity titre of 102.75 TCID50/ml). The results demonstrated that both the native breed and an improved pig breed are fully susceptible to CSFV infection
and the mortality rate is high. LWC pigs demonstrated lower (or shorter) survival times (50% survival time: 11 days), earlier
and higher pyrexia and earlier onset of viraemia compared to ML pigs (50% survival time: 18 days). In the context of village-based
pig production, the longer time from infection to death in native ML pigs means that incubating or early sick pigs are likely
to be sold once an outbreak of CSF is recognized in a village. This increased longevity probably contributes to the maintenance
and spread of disease in a population where generally the contact rate is low. 相似文献
18.
为配合猪瘟新型疫苗的研发,建立了猪瘟病毒NS3蛋白抗体检测间接ELISA,以期达到有效区分新型疫苗免疫猪与自然感染猪(包括常规疫苗接种猪)的目的。以接种猪瘟病毒(CSFV)石门株的PK-15细胞为模板提取总RNA,经特异性PCR扩增获得长度为2 049bp的CSFV NS3基因,将其克隆至插入了具有自聚集自切割功能短肽的原核表达载体pET-32a(+),在大肠埃希菌Rosetta(DE3)中优化表达CSFV石门株NS3基因。Western blot分析表明重组蛋白NS3具有反应原性。将纯化的重组蛋白NS3作为包被抗原建立检测CSFV NS3抗体的间接ELISA,以美国爱德士(Idexx)猪瘟病毒抗体检测试剂盒抗体检测结果为标准,对502份血清样品进行检测。结果表明,所建立方法的特异性为96.9%,敏感性为89.7%,总符合率为95.8%,为猪瘟新型疫苗的推广应用提供了血清学检测方法。 相似文献
19.
1. The significance of airborne transmission in epidemics of infectious diseases in the livestock production industry remains unclear. The study therefore investigated the shedding route (faeces vs. exhaled air) of a vaccine strain of infectious bursal disease virus (IBDV) by broilers and the emission of airborne virus. 2. The experimental room contained 526 broilers which were orally inoculated at the age of 20?d. The airborne virus was sampled by three different bioaerosol samplers: Andersen six-stage impactor, all-glass impinger (AGI-30) and OMNI-3000. 3. Infected broilers started to shed virus in faeces on d 5 post inoculation (PI), and stopped shedding on d 12 PI. The faecal virus remained detectable for at least two d after drying under broiler room conditions. No virus was detected in the air exhaled by broilers. 4. Airborne virus was collected on d 5, 8 and 12 PI at 20?cm above the floor, and on d 8 and 12 PI in exhausted air. The emission rates of IBDV were 4·0 log10 50% tissue culture infectious dose (TCID50)/bird/d on d 8 PI, and 4·5 log10 TCID50/bird/d on d 12 PI. 5. We concluded that broilers shed IBDV mainly through their faeces. The presence of indoor airborne virus is associated with the viral presence in faeces. The successful recovery of airborne virus in exhausted air indicates there is a potential risk of virus spreading to the ambient environment via air. 相似文献