贵州省安龙县狂犬病的流行病学研究

为了探讨局部地区暴发流行的因素,对2004年与2005年间在贵州省安龙县及其周边发生的人间狂犬病进行个案调查,用直接免疫荧光法检测犬脑组织中的狂犬病毒抗原,用RT-PCR法扩增G基因片段,测定核苷酸序列并构建系统发生树进行遗传特征分析.自1994-2003年安龙县报告无人间狂犬病疫情,但于2004与2005年分别报告44例与27例人狂犬病.71例狂犬病患者中68人被犬所伤,3人被猫所伤.在有完整记录的69例患者中,潜伏期平均为47.5d(10～282),13例狂犬病患者的潜伏期≤15d(18.84%);32例的潜伏期≤30d(46.38%),尤其是在2004年的患者中,潜伏期≤20d的病例有13例,占30%.2005年在疫点周围采集的85只犬脑组织中,5只犬脑组织狂犬病毒抗原阳性,阳性率为5.88%.A13株与A48株病毒之间的G基因核苷酸序列同源性为86.5%,与已报道病毒株比较发现,A13株与广西分离株有最高的同源性,而A48株却与印度尼西亚分离株SN01-23的同源性最高.比较分析两株病毒G基因编码的氨基酸序列,发现包括GⅠ、GⅡ,以及GⅢ抗原位点内的区间,A13与A48分别有19和28个氨基酸位点发生了氨基酸的替代.用全G基因核苷酸序列进行系统分析发现,两株毒株全为Ⅰ型狂犬病毒.结果表明引起安龙县的狂犬病流行的病毒不是新型狂犬病毒,犬饲养量大与病毒携带率高是狂犬病发病暴发流行的直接原因.     

贵州省安龙县狂犬病的流行病学研究

为了探讨局部地区暴发流行的因素,对2004年与2005年间在贵州省安龙县及其周边发生的人间狂犬病进行个案调查,用直接免疫荧光法检测犬脑组织中的狂犬病毒抗原,用RT-PCR法扩增G基因片段,测定核苷酸序列并构建系统发生树进行遗传特征分析。自1994-2003年安龙县报告无人间狂犬病疫情,但于2004与2005年分别报告44例与27例人狂犬病。71例狂犬病患者中68人被犬所伤,3人被猫所伤。在有完整记录的69例患者中,潜伏期平均为47.5d(10～282),13例狂犬病患者的潜伏期≤15d(18.84%);32例的潜伏期≤30d(46.38%),尤其是在2004年的患者中,潜伏期≤20d的病例有13例,占30%。2005年在疫点周围采集的85只犬脑组织中,5只犬脑组织狂犬病毒抗原阳性,阳性率为5.88%。A13株与A48株病毒之间的G基因核苷酸序列同源性为86.5%,与已报道病毒株比较发现,A13株与广西分离株有最高的同源性,而A48株却与印度尼西亚分离株SN01-23的同源性最高。比较分析两株病毒G基因编码的氨基酸序列,发现包括GⅠ、GⅡ,以及GⅢ抗原位点内的区间,A13与A48分别有19和28个氨基酸位点发生了氨基酸的替代。用全G基因核苷酸序列进行系统分析发现,两株毒株全为Ⅰ型狂犬病毒。结果表明引起安龙县的狂犬病流行的病毒不是新型狂犬病毒,犬饲养量大与病毒携带率高是狂犬病发病暴发流行的直接原因。     

用Vero细胞制备马抗狂犬病血清抗原

以Vero细胞为基质制备马抗狂犬病血清用抗原,以期建立有效、经济、简便的抗原制备方法.用含10%马血清营养液对Vero细胞作适应性培养,接种狂犬病毒,以含1%～3%马血清营养液作维持液培养病毒,于第5,8天收获病毒液,经灭活、浓缩、离心等制成抗原.第5,8天收获病毒滴度可稳定在7.0logLD50/mL以上,灭活抗原具良好的抗原性,用NIH法测定效价达6.0IU/mL以上,可用作抗原生产抗狂犬病血清.     

抗狂犬病毒中和抗体研究进展

狂犬病是一种人兽共患传染病,人和动物一旦发病后死亡率几乎百分之百,而有效的暴露后预防措施可以将死亡风险降至零。根据WHO推荐的狂犬病暴露后预防方案,一般狂犬病暴露者需要进行疫苗注射,严重者则需在进行疫苗注射的同时注射抗狂犬病毒中和抗体。常用的中和抗体有马抗狂犬病毒免疫球蛋白和人抗狂犬病毒免疫球蛋白,然而两者都存在引起过敏反应或血液疾病的风险。人源抗狂犬病毒中和抗体则因为具有安全性高、成本低、可量产等优点有望代替免疫球蛋白用于暴露后预防。基因工程抗体技术的发展加速了抗体人源化的进程。就抗狂犬病毒中和抗体的发展历程,不同类型中和抗体的优缺点以及中和抗体的未来研究方向作了综述及展望,以期为新一代狂犬疫苗的研发提供参考。     

表达绿色荧光蛋白嵌合狂犬病病毒HEP-GFP株的拯救

狂犬病毒(Rabies Virus,RV)是人和犬、猫等多种动物狂犬病的病原,其基因组是单股负链RNA,基因组结构为3'-核蛋白(N)基因-磷蛋白(P)基因-基质蛋白(M)基因-糖蛋白(G)基因-大蛋白(L)基因-5'.     

伪狂犬病毒固有免疫逃避机制

伪狂犬病毒能引起多种动物伪狂犬病,严重威胁养殖业发展。尤其自2011年新毒株在我国的大面积流行,使伪狂犬病的防控研究再次成为兽医学和病毒学的热点。其中,伪狂犬病毒介导的固有免疫逃避机制是其防控困难的重要原因,对于这一问题的探讨将有助于新疫苗和特效治疗药物的研发。因此,本文旨在对伪狂犬病毒固有免疫逃避机制的研究进展进行综述,为伪狂犬病的有效防控提供理论指导。     

用Vero细胞制备马抗狂犬病血清抗原

以Vero细胞为基质制备马抗狂犬病血清用抗原,以期建立有效、经济、简便的抗原制备方法。用含10％马血清营养液对Vero细胞作适应性培养,接种狂犬病毒,以含1％～3％马血清营养液作维持液培养病毒,于第5,8天收获病毒液,经灭活、浓缩、离心等制成抗原。第5,8天收获病毒滴度可稳定在7．010gLD50／mL以上,灭活抗原具良好的抗原性,用NIH法测定效价达6．0IU／mL以上,可用作抗原生产抗狂犬病血清。     

狂犬病毒糖蛋白综述

狂犬病是一种由狂犬病毒引起的高度致命的人兽共患传染病,在五种狂犬病毒蛋白中,糖蛋白是唯一暴露在包膜外的,是一种保护性抗原,可以有效地诱导机体产生中和性抗体。本文主要重点介绍了糖蛋白的结构与功能。     

狂犬病病毒抗体胶体金检测试纸的制备

通过胶体金免疫层析技术建立一种特异、便捷、快速的狂犬病病毒抗体检测方法,对犬等动物免疫狂犬病疫苗后的抗体水平监测提供参考。用醋酸锌沉淀法沉淀狂犬病病毒CVS11,Sepharose 4FF进行层析纯化。用柠檬酸三钠还原法制备的胶体金,标记纯化的狂犬病病毒,喷于试纸的结合释放垫 (金标垫),将SPA (葡萄球菌表面A蛋白) 和纯化的兔抗狂犬病病毒IgG分别喷于试纸的T (检测线) 处和C (对照线) 处,组装试纸条。用制备的试纸条对261份犬血清进行检测,与快速荧光灶抑制试验 (RFFIT) 检测的结果一致;对已知效价的犬狂犬病毒中和抗体 (VNA) 大于0.5 IU,结果为阳性;对狂犬病病毒中和抗体 (VNA) 小于0.5 IU/mL的血清,结果为阴性。制备的狂犬病病毒抗体胶体金检测试纸检测犬血清抗体,具有特异、便捷、快速的特点,能够检测出狂犬病病毒中和抗体大于0.5 IU的血清,适用于临床犬血清抗体水平监测,具有良好的应用前景。     

病毒感染复数对狂犬病毒PM株增殖的影响

目的探讨不同感染复数(Multiplicity of infection,MOI)的狂犬病毒PM株在人二倍体细胞(2BS株)中增殖的影响,确定感染复数。方法将狂犬病毒PM株按照MOI 0.01、0.02、0.05、0.10、0.15、0.20接种至2BS细胞中,用显微镜观察接种病毒后细胞的形态变化;培养3~5 d后,第一次收获病毒液;更换培养液继续培养3~4 d后,进行第二次收获。采用小鼠脑内滴定法测定狂犬病毒滴度,通过NIH法测定灭活病毒液的效价。结果当MOI为0.05~0.10时,细胞圆缩30%~40%,细胞能维持较好的生长形态,可收获病毒,病毒收获液的滴度较高(5.0lg LD50/m L),且灭活后病毒液的效价较高(4.0IU/m L)。结论确定狂犬病毒PM株在人二倍体2BS细胞增殖的适宜MOI,为人用狂犬病疫苗生产工艺的优化提供了依据。     

ELECTRON MICROSCOPE STUDIES OF RABIES VIRUS IN MOUSE BRAIN

The cells of brains of 2- and 3-day old mice infected with street rabies virus were examined in the electron microscope. It was observed that characteristic rod-like or elongated particles were found within a "matrix" in the cytoplasm of nerve cells and of astrocytes. These rod-like particles can be separated into two types, on the basis of slightly different morphological features. One particle is 110 to 120 mµ wide and has double-membraned coats; the other is 120 to 130 mµ wide and is covered by a single limiting membrane. The former is closely associated with the endoplasmic reticulum. The biological relationship between the two types is unknown, but both types of particles are considered to be street rabies viruses because of their structural features. It is believed that segmentation and branching of elongated particles may play a role in virus multiplication. Negri bodies appear as dense round bodies containing various coarse structures but no virus particles.     

几种药物抑制马传染性贫血病毒和人免疫缺陷病毒的实验研究

用马传染性贫血病毒—驴胚肺二倍体细胞(EIAV-DELDC)为实验体系,以细胞中病毒逆转录酶活性及病毒相关抗原的表达为观察指标,检测了叠氮胸苷(AZT)、三氮唑核苷(Ribavirin,病毒唑)、磷羧基甲酸钠(PFA)和苏拉明等4种已知抗人免疫缺陷病毒(HIV-1)药物对马传染性贫血病毒的抑制作用。结果表明,PFA、AZTTP(三磷酸AZT)和苏拉明均能抑制病毒相关抗原的表达,AZT虽无此作用,但能抑制细胞内逆转录酶活性。用~3H-TMP掺入法比较了PFA、AZTTP、苏拉明对体外无细胞系EIAV逆转录酶粗提物和HIV-1基因工程产物逆转录酶活性的抑制作用表明,两种逆转录酶对苏拉明的敏感性相近,而HIV-1逆转录酶对PFA和AZTTP的敏感性较EIAV者高约100倍。又以无细胞系中逆转录酶活性测定法,检测了12种中药提取物的抑制作用,其中小柴胡汤对EIAV和HIV-1逆转录酶活性都有抑制作用,IC_(50)为717μg/ml和700μg/ml(生药浓度)。小柴胡汤对两种病毒感染细胞中抗原的表达和HIV引起细胞病变都有抑制作用,对HIV-1的抑制比EIAV强。这些结果表明,EIAV-DELDC体系可考虑作为抗HIV-1药物筛选模型。     

家蚕核型多角体病毒对小鼠的易感性及其蛋白组份的检测

家蚕核型多角体病毒(Bm-NPV)是一类环状DNA病毒。该病毒作为载体,应用在杆状病毒-昆虫真核生物表达系统。为确定家蚕基因工程表达系统产品对人的安全性,观察了该病毒对哺乳动物细胞株及对小鼠的易感性,建立了检测Bm-NPV蛋白组份的dot-ELISA方法,可用于生物表达器生产的产品中残存多角体病毒组份的检测。结果表明,芽生型多角体病毒(BV)在杂交瘤细胞和HL60细胞中不增殖。不同剂量包埋型多角体病毒(OV)灌胃感染小鼠,小鼠肝、肾组织切片的电镜观察及免疫组化染色未见到病毒颗粒。     

THE LINE-PATTERN VIRUS DISEASE OF PLUMS

The disease, now usually called line-pattern of plum, has been described under many names in most countries where plums are grown extensively.
Naturally infected trees show widely differing Symptoms; this has two causes: (1) different varieties react differently to the same isolate of the virus, and (2) different isolates cause different symptoms in the same variety. Because the virus occurs in strains with different pathogenicities, the choice of indicator varieties is important when selecting virus-free material by transmission tests. Peach seedlings and the mazzard clone, F 12/1, were the most sensitive types found.
The line-pattern virus does not become fully systemic in some varieties of plum. In this and other respects, it resembles the viruses that cause apple mosaic; three isolates obtained from plum and two from apple produced similar Symptoms in peach and apple. It is therefore suggested that plum line-pattern and apple mosaic viruses are caused by strains of one virus.     

VIRUS DISEASES OF CACAO IN WEST AFRICA IX. STRAIN VARIATION AND INTERFERENCE IN VIRUS 1A

Cacao virus iA, the most important and prevalent of the viruses that attack cacao in the Gold Coast, occurs in strains that differ widely in their virulence towards cacao. Outbreaks usually contain trees infected with different strains and individual trees are often infected simultaneously with more than one strain; this can be demonstrated by coppicing the trees, and by inoculating sets of test plants with grafts from different parts of one tree. Neither mild nor virulent strains seemed to be consistently dominant in roots or in other parts of cacao trees.
Cacao plants infected with mild strains were nearly always protected against the effects of infection by virulent strains; however, virulent strains entered hosts already infected with mild strains, but usually without causing any symptoms unless the plants were coppiced. The severe symptoms that developed on new growth from such coppiced plants were seldom repeated in later growth. Mealybugs transmitted the virulent strains from leaves with symptoms characteristic of infection by the latter, but not from leaves free from such symptoms. These results suggest that the multiplication of a virulent strain is impeded in plants infected with a mild strain.
In the field, infection with a mild strain protected mature trees against the effects of virulent strains spread by mealybugs. During 3 years in which 273 out of 387 previously uninfected trees became severely diseased, only 35 out of 416 infected with themildstrain developed symptoms of infectionwiththevirulent strain. Five years after infection with the mild strain, trees were yielding I pod per tree more than in the year they were infected, whereas the decrease on trees infected with the virulent strain was 16 pods per tree. Some limitations in the practical application of protection by mild strains, and objections to its use as a control measure, are discussed.