关键词： 儿童预防接种   护理干预   系统   接种百白破疫苗   脊髓灰质炎疫苗   预防接种率   群众基础   接种疫苗  

摘要： 预防接种是一项投资少、效益高并具有广泛群众基础的疾病防控措施。预防接种旨在通过接种疫苗，预防相应的传染病，它是一种最有效、最方便、最经济的手段。2010年6月至2011年7月，我院体检保健中心共为2542名儿童接种百白破疫苗、脊髓灰质炎疫苗、麻疹疫苗、卡介苗、乙肝疫苗和流感疫苗等，并对其在接种前、中、后进行系统的护理干预，均取得了良好的效果，保证儿童预防接种率达到国家要求的标准，现报告如下。

关键词： 脊髓灰质炎病毒   D抗原   C抗原   抗体   单克隆   酶联免疫吸附测定  

摘要： 目的制备脊髓灰质炎病毒(Poliovirus)Ⅰ型D抗原单克隆抗体,并建立双抗体夹心ELISA检测方法。方法采用Vero细胞培养脊髓灰质炎病毒Ⅰ型,柱层析纯化病毒抗原,SDS-PAGE鉴定病毒蛋白,HPLC分析抗原纯度,Western blot分析抗原的反应原性。以纯化的脊髓灰质炎病毒Ⅰ型D抗原免疫BALB/c小鼠,进行细胞融合,筛选可分泌脊髓灰质炎病毒I型D抗原单克隆抗体的杂交瘤细胞株。单克隆抗体经纯化后,采用间接ELISA法检测小鼠腹水抗体效价、单抗的特异性及相对亲和力。分别以纯化的单抗作为捕获抗体和酶标抗体,进行配对试验,建立用于脊髓灰质炎病毒Ⅰ型D抗原检测的双抗体夹心ELISA方法,确定方法的最佳线性范围,并验证方法的特异性。结果纯化的脊髓灰质炎病毒Ⅰ型D抗原经SDS-PAGE分析,共显示4条蛋白条带,相对分子质量分别为约40 000、35 000、32 000和28 000;经HLPC分析,纯度达99%;小鼠免疫血清可与相对分子质量约为35 000的VP1蛋白发生特异性反应。共筛选出5株可稳定分泌脊髓灰质炎病毒I型D抗原单克隆抗体的阳性杂交瘤细胞株,纯化的单抗仅与D抗原发生反应;2H1和3B5株单抗的效价分别为2×107和2×106,且亲和力高于其他3株单抗。采用2H1和3B5株单抗配对,可以特异性检测D抗原,而与C抗原不反应;D抗原最佳线性范围为4.300～0.071 DU/ml,R2为0.994 6。该法仅可检出脊髓灰质炎病毒Ⅰ型D抗原,而与脊髓灰质炎病毒Ⅱ型、Ⅲ型D抗原和脊髓灰质炎病毒Ⅰ型C抗原、EV71抗原、Vero细胞培养上清不发生交叉反应。结论制备了脊髓灰质炎病毒Ⅰ型D抗原单克隆抗体,建立了可用于脊髓灰质炎病毒Ⅰ型D抗原特异性检测的双抗体夹心ELISA方法。

关键词： POLIOVIRUS   VIRAL replication   VIRAL genetics   CELL membranes   CELL morphology   VIRUSES -- Morphology  

摘要： Replication of all positive-strand RNA viruses is intimately associated with membranes. Here we utilize electron tomography and other methods to investigate the remodeling of membranes in poliovirus-infected cells. We found that the viral replication structures previously described as "vesicles" are in fact convoluted, branching chambers with complex and dynamic morphology. They are likely to originate from cis-Golgi membranes and are represented during the early stages of infection by single-walled connecting and branching tubular compartments. These early viral organelles gradually transform into double-membrane structures by extension of membranous walls and/or collapsing of the luminal cavity of the single-membrane structures. As the double-membrane regions develop, they enclose cytoplasmic material. At this stage, a continuous membranous structure may have double-and single-walled membrane morphology at adjacent cross-sections. In the late stages of the replication cycle, the structures are represented mostly by double-membrane vesicles. Viral replication proteins, double-stranded RNA species, and actively replicating RNA are associated with both double-and single-membrane structures. However, the exponential phase of viral RNA synthesis occurs when single-membrane formations are predominant in the cell. It has been shown previously that replication complexes of some other positive-strand RNA viruses form on membrane invaginations, which result from negative membrane curvature. Our data show that the remodeling of cellular membranes in poliovirus-infected cells produces structures with positive curvature of membranes. Thus, it is likely that there is a fundamental divergence in the requirements for the supporting cellular membrane-shaping machinery among different groups of positive-strand RNA viruses.

关键词： VIRAL genetics   POLIOVIRUS   ANTIVIRAL agents   TOLL-like receptors   TRANSGENIC mice   RNA  

摘要： RIG-I-like receptors and Toll-like receptors (TLRs) play important roles in the recognition of viral infections. However, how these molecules contribute to the defense against poliovirus (PV) infection remains unclear. We characterized the roles of these sensors in PV infection in transgenic mice expressing the PV receptor. We observed that alpha/beta interferon (IFN-alpha/beta) production in response to PV infection occurred in an MDA5-dependent but RIG-I-independent manner in primary cultured kidney cells in vitro. These results suggest that, similar to the RNA of other picornaviruses, PV RNA is recognized by MDA5. However, serum IFN-alpha levels, the viral load in nonneural tissues, and mortality rates did not differ significantly between MDA5-deficient mice and wild-type mice. In contrast, we observed that serum IFN production was abrogated and that the viral load in nonneural tissues and mortality rates were both markedly higher in TIR domain-containing adaptor-inducing IFN-beta (TRIF)-deficient and TLR3-deficient mice than in wild-type mice. The mortality rate of MyD88-deficient mice was slightly higher than that of wild-type mice. These results suggest that multiple pathways are involved in the antiviral response in mice and that the TLR3-TRIF-mediated signaling pathway plays an essential role in the antiviral response against PV infection.

关键词： Infectious disease epidemiology   polio   polio vaccine virus   public health   surveillance  

摘要： Environmental poliovirus surveillance (ENV) means monitoring of poliovirus (PV) transmission in human populations by examining environmental specimens supposedly contaminated by human faeces. The rationale is based on the fact that PV-infected individuals, whether presenting with disease symptoms or not, shed large amounts of PV in the faeces for several weeks. As the morbidity: infection ratio of PV infection is very low, this fact contributes to the sensitivity of ENV which under optimal conditions can be better than that of the standard acute flaccid paralysis (AFP) surveillance. The World Health Organization has included ENV in the new Strategic Plan of the Global Polio Eradication Initiative for years 2010-2012 to be increasingly used in PV surveillance, supplementing AFP surveillance. In this paper we review the feasibility of using ENV to monitor wild PV and vaccine-derived PV circulation in human populations, based on global experiences in defined epidemiological situations.

关键词： POLIOVIRUS   HEALTH of refugees   HEALTH of immigrants   ENTEROVIRUS diseases   COXSACKIEVIRUS diseases   DISEASE prevalence  

摘要： The aim of this study was to evaluate the presence of wild poliovirus or sabin-like poliovirus in 152 stool samples from migrants in the Accommodation Center in Italy and liquid waste from the sewage systems. Two surveys were performed in 2008 and 2011. All stool samples were negative for enterovirus and poliovirus. One of the liquid samples analyzed was positive for Coxsackievirus type B5.

关键词： 脊髓灰质炎病毒   精神病状态评定量表   抗体   单克隆   基因连锁   酶联免疫吸附测定   敏感性与特异性   血型抗原   杂交瘤   动物   小鼠  

摘要： 目的:制备脊髓灰质炎病毒(Poliovims)I型D抗原单克隆抗体,并建立双抗体夹心ELISA检测方法。方法:采用Vero细胞培养脊髓灰质炎病毒I型,柱层析纯化病毒抗原,***鉴定病毒蛋白,HPLC分析抗原纯度,Westernblot分析抗原的反应原性。以纯化的脊髓灰质炎病毒I型D抗原免疫BALB/e小鼠,进行细胞融合,筛选可分泌脊髓灰质炎病毒I型D抗原单克隆抗体的杂交瘤细胞株。单克隆抗体经纯化后,采用间接ELISA法检测小鼠腹水抗体效价、单抗的特异性及相对亲和力。分别以纯化的单抗作为捕获抗体和酶标抗体,进行配对试验,建立用于脊髓灰质炎病毒I型D抗原检测的双抗体夹心ELISA方法,确定方法的最佳线性范围,并验证方法的特异性。结果:纯化的脊髓灰质炎病毒I型D抗原经***分析,共显示4条蛋白条带,相对分子质量分别为约40000、35000、32000和28000;经HLPC分析,纯度达99%;小鼠免疫血清可与相对分子质量约为35000的VPl蛋白发生特异性反应。共筛选出5株可稳定分泌脊髓灰质炎病毒I型D抗原单克隆抗体的阳性杂交瘤细胞株,纯化的单抗仅与D抗原发生反应;2H1和385株单抗的效价分别为2×10'和2×106,且亲和力高于其他3株单抗。采用2H1和385株单抗配对,可以特异性检测D抗原,而与C抗原不反应;D抗原最佳线性范围为4.300—0.071DU/ml,R:为0.9946。该法仅可检出脊髓灰质炎病毒I型D抗原,而与脊髓灰质炎病毒Ⅱ型、Ⅲ型D抗原和脊髓灰质炎病毒I型C抗原、EV71抗原、ver0细胞培养上清不发生交叉反应。结论:制备了脊髓灰质炎病毒I型D抗原单克隆抗体,建立了可用于脊髓灰质炎病毒I型D抗原特异性检测的双抗体夹心ELISA方法。

关键词： hormesis   biphasic   hormetic   polio   ethanol   Salk vaccine  

摘要： The production of the Salk vaccine polio virus by monkey kidney cells was generated using the synthetic tissue culture medium, Mixture 199. In this paper's retrospective assessment of this process, it was discovered that Mixture 199 was modified by the addition of ethanol to optimize animal cell survival based on experimentation that revealed a hormetic-like biphasic response relationship. This hormesis-based optimization procedure was then applied to all uses of Mixture 199 and modifications of it, including its application to the Salk polio vaccine during preliminary testing and in its subsequent major societal treatment programs.

关键词： Poliovirus   Reovirus   Interference   Coinfection  

摘要： Objective: Viral interference has been demonstrated in different systems, such as the effect of enterovirus infection on live-attenuated oral polio vaccine. In this study, the effect of reovirus which could exist in the human intestinal tract on poliovirus vaccine strains was investigated and could be an important factor to consider in oral polio vaccination. Methods: Cells were infected with reovirus, then superinfected with poliovirus. The amount of viral yields was measured by the TCID(50) and plaque assay methods. Polioviral RNA synthesis was studied by real-time RT-PCR and the viral RNA load was calculated. Viral protein synthesis was determined using the techniques of immunoflourescent staining and PAGE followed by the immunoblotting experiment. Results: Poliovirus superinfection of reovirus-infected cells resulted in inhibition of poliovirus replication. It was found that the inhibitory effect of reovirus was after establishment of its infection (2 h postinfection). There was no competition between the two viruses for cell attachment but poliovirus RNA and protein synthesis were inhibited. Conclusion: Infection of cells with reovirus could interfere with the growth of poliovirus upon superinfection. This phenomenon could be important to consider when using attenuated poliovirus vaccine. Copyright (C) 2011 S. Karger AG, Basel

关键词： 疫苗衍生脊髓灰质炎病毒   免疫缺陷   口服脊髓灰质炎减毒活疫苗  

摘要： 目的:分析1例免疫缺陷患儿排出Ⅱ型疫苗衍生脊髓灰质炎病毒病原学检测结果及对当地维持无脊灰的影响。方法:对免疫缺陷患儿粪便标本及其密切接触者粪便标本和污水标本采用病毒分离的方法,对分离到的Ⅱ型脊灰病毒进行VP1编码区基因核苷酸序列测定和分析。结果:2011年在贵州省1例AFP病例双份粪便标本中,分离到2株Ⅱ型脊灰病毒(PV),经核苷酸序列测定和分析鉴定为Ⅱ型VDPVs,临床诊断为原发性免疫缺陷病,患儿曾接种过4剂口服脊灰减毒活疫苗(OPV)。对该患儿连续采集粪便标本检测,未再发现VDPV。贵州省AFP病例分类专家诊断小组诊断为周期性麻痹、VDPV携带者。结论:此次发现的Ⅱ型VDPV发生在OPV高免疫覆盖率地区,未发现形成VDPV循环的证据。